MX338563B

Antagonists of il-6 to prevent or treat cachexia, weakness, fatigue and/or fever.

Abstract

The present invention is directed to therapeutic methods using antibodies and fragments thereof having binding specificity for IL-6 to prevent or treat cachexia, fever, weakness and/or fatigue in a patient in need thereof. In preferred embodiments, the anti- IL-6 antibodies will be humanized and/or will be aglycosylated. Also, in preferred embodiments these patients will comprise those exhibiting (or at risk of developing) an elevated serum C-reactive protein level. In another preferred embodiment, the patient's survivability or quality of life will preferably be improved.

MX338563B, drawing sheet 1 — Sheet 1 of 743

Term

3.2 yearsleft in the term

Expires 24 November 2029.

35 claims: 7 independent, 28 dependent

1
CLAIMS REIVINDICACIONES IMPI IMPI 1. El uso de un anticuerpo o fragmento de anticuerpo anti-IL-6 para la manufactura de un medicamento para tratar anemia en un paciente que tiene un desorden que envuelve elevada interleucina-6 (IL-6), en donde el anticuerpo o fragmento de anticuerpo anti-IL-6 comprende una secuencia región determinante de la complementariedad (CDR)1 de cadena ligera variable de SEQ ID NO:4, secuencia CDR2 de SEQ ID NO:5 y secuencia CDR3 de SEQ ID NO: 6, y una secuencia CDR1 de cadena pesada variable de SEQ ID NO:7, secuencia CDR2 de SEQ ID NO:8 o one. The use of an anti-IL-6 antibody or antibody fragment for the manufacture of a medicament to treat anemia in a patient who has an elevated interleukin-6 (IL-6) enveloping disorder, wherein the antibody or antibody fragment anti-IL-6 comprises a variable light chain complementarity determining region (CDR) 1 sequence of SEQ ID NO: 4, CDR2 sequence of SEQ ID NO: 5 and CDR3 sequence of SEQ ID NO: 6, and a variable heavy chain CDR1 sequence of SEQ ID NO: 7, CDR2 sequence of SEQ ID NO: 8 or SEQ ID NO: 120, and CDR3 sequence of SEQ ID NO: 9. SEQ ID NO:120, y secuencia CDR3 de SEQ ID NO:9.
4
4. El uso de conformidad con la ΐΝ,ίτσντν wí-.'rrxso oe (Μ$7<Μ en donde '»ma» reivindicación 1, el anticuerpo o fragmento de anticuerpo anti-IL-6 comprende el polipéptido de cadena pesada variable que tiene la secuencia de aminoácidos mostrada en la SEQ ID NO:657 y el polipéptido de cadena ligera variable que tiene la secuencia de aminoácidos mostrada en la SEQ ID NO: 709. Using in accordance with ΐΝ, ίτσντν wí -. 'Rrxso oe (Μ $ 7 <Μ where' 'ma' 'claim 1, the anti-IL-6 antibody or antibody fragment comprises the variable heavy chain polypeptide having the amino acid sequence shown in SEQ ID NO: 657 and the variable light chain polypeptide having the amino acid sequence shown in SEQ ID NO: 709.
16
16. donde el where he
17
17. donde el una vida días. where the one day life.
18
18. donde el una vida días. where the one day life.
19
19. donde el una vida días. where the one day life. El uso de conformidad con la reivindicación 15, en The use according to claim 15, in Fc Human comes from IgGl, IgG2, IgG3 or IgG4. Fc humano proviene de IgGl, IgG2, IgG3 o IgG4. El uso de conformidad con la reivindicación 1, en anticuerpo o fragmento de anticuerpo anti-IL-6 tiene media de eliminación de al menos aproximadamente 22 The use according to claim 1, in anti-IL-6 antibody or antibody fragment has elimination average of at least about 22 El uso de conformidad con la reivindicación 1, en anticuerpo o fragmento de anticuerpo anti-IL-6 tiene media de eliminación de al menos aproximadamente 25 The use according to claim 1, in anti-IL-6 antibody or antibody fragment has elimination average of at least about 25 El uso de conformidad con la reivindicación 1, en anticuerpo o fragmento de anticuerpo anti-IL-6 tiene media de eliminación de al menos aproximadamente 30 The use according to claim 1, in anti-IL-6 antibody or antibody fragment has elimination average of at least about 30
25
A pharmaceutical composition comprising a 25. Una composición farmacéutica que comprende un 633 ¡«IfTeTijTO M6TOCANO O £ L * AJOMEDAO 633 ¡«IfTeTijTO M6TOCANO O£ L* AJOMEDAO ΙΪΜΧβΠϋΛΓ anti-IL-6 antibody or antibody fragment comprising a variable light chain CDRl sequence of SEQ ID NO:4, CDR2 sequence of SEQ ID NO: 5, and CDR3 sequence of SEQ ID NO: 6, and a CDRl of SEQ ID NO: 7 Variable Heavy Chain, Sequence ΙΪΜΧβΠϋΛΓ anticuerpo o fragmento de anticuerpo anti-IL-6 que comprende una secuencia CDRl de cadena ligera variable de SEQ ID NO: 4, secuencia CDR2 de SEQ ID NO:5, y secuencia CDR3 de SEQ ID NO:6, y una CDRl de cadena pesada varible de SEQ ID NO:7, secuencia CDR2 de SEQ ID NO:8 o SEQ ID NO:120, y secuencia CDR3 de SEQ ID CDR2 of SEQ ID NO: 8 or SEQ ID NO: 120, and CDR3 sequence of SEQ ID NO: 9, for use in treating anemia in a patient having a disorder associated with elevated interleukin-6, wherein the antibody or antibody fragment is adapted to be administrable to the patient in a dosage regimen comprising a or more doses of approximately NO: 9, para usarse en el tratamiento de la anemia que tiene un paciente que tiene un desorden asociado con interleucina-6 elevada, en donde el anticuerpo o fragmento de anticuerpo esta adaptado para ser administrable al paciente en un régimen de dosificación que comprende una o más dosis de aproximadamente 80 mg, about 160 mg or about 320 mg. 80 mg, aproximadamente 160 mg o aproximadamente 320 mg. Antibody or antibody fragment is contained in phosphate buffered saline. anticuerpo o fragmento de anticuerpo está contenida en solución salina amortiguada con fosfato.

Independent claims7

4,809 paragraphs in 3,567 sections, as filed

(54) Title: IL-6 ANTAGONISTS TO PREVENT OR TREAT CAQUEXIA, WEAKNESS, FATIGUE AND / OR FEVER.

(54) Title: ANTAGONISTS OF IL-6 TO PREVENT OR TREAT CACHEXIA, WEAKNESS, FATIGUE AND / OR FEVER.

(57) Summary

The present invention relates to the use of an anti-IL6 antibody or antibody fragment for the manufacture of a medicament for treating anemia in a patient having an elevated interleukin-6 (IL-6) enveloping disorder, wherein the antibody or anti-IL-6 antibody fragment comprises a variable light chain complementarity determining region (CDR) 1 sequence of SEO ID NO: 4, CDR2 sequence of SEO ID NO: 5 and CDR3 sequence of SEQ ID NO: 6, and a variable heavy chain CDR1 sequence of SEQ ID NO; 7, CDR2 sequence of SEQ ID NO: 8 or SEQ ID NO: 120, and CDR3 sequence of SEQ ID NO: 9. A pharmaceutical composition comprising an anti-IL-6 antibody or antibody fragment comprising a variable light chain CDR1 sequence of SEQ ID NO: 4, CDR2 sequence of SEQ ID NO: 5, and CDR3 sequence of SEQ ID NO: 6 , AND A variable heavy chain CDR1 of SEQ ID NO: 7, CDR2 sequence of SEQ ID No: 8 or SEQ ID NO: 120, and sequence of SEQ ID NO: 9, for use in treating anemia in a patient having an elevated interleukin-6 associated disorder, wherein the antibody or antibody fragment is adapted to be administrable to the patient in a dosage regimen comprising one or more doses of about 80 mg, about 160 mg or about 320 mg.

(57) Abstract

The present invention is directed to therapeutic methods using antibodies and fragments thereof having binding specificity for IL-6 to prevent or treat cachexia, fever, weakness and / or fatigue in a patient in need thereof. In preferred embodiments, the anti- IL-6 antibodies will be humanized and / or will be aglycosylated. Also, in preferred performances these patients will comprise those exhibiting (or at risk of developing) an elevated serum C-reactive protein level. In another preferred embodiment, the patient's survivability or quality of Ufe will preferably be improved.

Institute

Mexican Property

Industrial _SE «ΗΠΆΚίΑ I heard: HWaK ίΐΒββ '\ í»? ..or

PATENT TITLE MOk338563

Owner (s): ALDERBIO HOLDINGS LLC. -

Address: 101 Convention Center Drive, Suite 850, Las Vegas, Nevada, 89109, USA

Name: IL-6 ANTAGONISTS TO PREVENT OR TREAT CAQUEXIA, WEAKNESS, FATIGUE AND / OR FEVER.

Classification: lnt.CI.8: A61K38 / 20; A61K39 / 395; A61P37 / 06; C07K16 / 24

Inventor (s): JEFFREY TL SMITH; RANDALL C. SCHATZMAN; MARK LITTON; JOHN

LATHAM

Number:

MX / a / 2011/005406

8OLICI

Date i

CITUD of presentation i of November 2009

PRIORITY

<td>Country: á</td><td>Date:</td><td></td><td>Number:</td>

<td>US</td><td>November 25, 2008</td><td></td><td> 61/117,811</td>

<td>US</td><td>November 25, 2008</td><td></td><td> 61/117,861</td>

<td>US</td><td>November 25, 2009.</td><td></td><td> 61/117,839</td>

<td>US</td><td>February 5, 2009</td><td></td><td> 12/366,567</td>

<td>US</td><td>February 24, 2009 i, · i</td><td rowspan="2">tea?</td><td> 12/391,717</td>

<td>US</td><td>February 24, 2009</td><td> 12/391,615</td>

<td>US</td><td>March 6, 2009 ”'</td><td>4Kt '' t 1</td><td> 12/399,156</td>

Validity: Twenty years

Expiration Date: November 24, 2029

The reference patent is granted jointly in articles 1 *, 2 * section V, 6 section III, and 59 of the Industrial Property Law.

In accordance with article 23 of Counted as of the date of presi rights. h

I Industrial Property Law. The present patent has a validity of twenty irreplaceable years, the international application is pending and subject to the payment of the fee to keep the

Whoever subscribes to this title does so based on the provisions of articles 6 fractions III and 7 bis 2 of the Official Industrial Property Law XDiarSS cfia Eadacacián ^ O.QJ ·) 27 / P6 / 1991 ,. amended on 05/02/1934, 10/25/1996,, 12/26/1997, 13 ^) 5/1999, 01/26/2004, 16, Ό6 / 2035, 01/25/2006, oOTKBoWOTÍ / 2010, 06/18/2010, 06/28/201 OR, 04/27/2012 and 04/09/2012); Articles '1 *, 3' section V subsection a), 4th and 12th sections I and III of the Regulations of the Mexican Institute of Industrial Property (DOF 12/14/1999, amended on 07/01/2002, 07/15/2004, 07/28/2004 and 09/07/2007); articles 1, 3, 4, 5 fraction V Clause a), 16 fractions I and III and 30 of the Organic Statute of the Mexican Institute of Industrial Property (DOF 12/27/1999, amended on 10/10/2002, 07/29/2004, 08/04/2004 and 09/13/2007); 1, 3 and 5 subsection a) of the Agreement that delegates powers to the Deputy Directors General, Coordinator, Divisional Directors, Heads of Regional Offices, Divisional Deputy Directors, Departmental Coordinators and other subordinates of the Mexican Property Institute (DOF 15 / 12/1999, amended on 02/04/2000, 07/29/2004, 08/04/2004 and 09/13/2007).

Issue Date: April 21, 2016

THE DIVISIONAL DIRECTOR OF PATENTS

NAHANNY CANAL REYES

Arenal No. 550, Floor 1.

Col. Pueblo Santa María Tepapan, Xochimtlco, CP 16020,

Mexico City

Tel {551 53 34 0 ~ 00 www.impi sob.tnx

MX / 2016/30977

20// /&//)&.

IL-6 ANTAGONISTS TO PREVENT OR TREAT CAQUEXIA,

WEAKNESS, FATIGUE AND / OR FEVER

CROSS REFERENCE with RELACJO REQUESTS ^ pj ^ j c MEXICAN INSTITUTE

J OF PROPERTY J

INDUSTRIAL >

This application claims the priority benefit of US provisional patent application No. 61 / 117,811, 61 / 117,861, and 61 / 117,839 all filed on November 25, 2008; and it is also a continuation in part of US application 12 / 502,581 filed on July 14, 2009, US serial number 12/399, 156 filed on March 6, 2009, US serial number 12 / 391,717 filed on February 24, 2009 and No. US Serial No. 12 / 366,567 filed February 5, 2009, the disclosure of which is incorporated herein by reference in its entirety.

The sequence listing in the file named

67858o707002.txt with a size of 332,004 bits that was created on November 24, 2009 is incorporated herein in its entirety by reference.

BACKGROUND OF THE INVENTION

Field of the Invention

The present invention is an extension of an earlier invention of the applicants described in the above-referenced patent applications relating to antibodies

INÍT1 MEXICAN TUTO , NS n £ INDUSTRIAL PROPERTY --- novel anti-IL-6 and novel therapies and protocols. therapeutics using anti-IL-6 antibodies, preferably those described herein. In particular, this invention relates to methods of preventing or treating cachexia, weakness, fatigue and / or fever in a patient in need, comprising administering to the patient an anti-IL-6 antibody or antibody fragment, whereby the cachexia, weakness, fatigue and / or fever of the patient are improved.

In another embodiment, this invention relates to methods of preventing or treating cachexia, weakness, fatigue, and / or fever in a patient in need, comprising administering to the patient an anti-IL-6 antibody or antibody fragment. , thereby improving the cachexia, weakness, fatigue and / or fever of the patient and monitoring the patient to assess cachexia, weakness, fatigue and / or fever, wherein the anti-IL-6 antibody or antibody fragment specifically binds to the same linear or conformational epitope (s) and / or competes to bind to the same epitope (s) s) linear (s) or conformational (s) in an intact human IL-6 polypeptide or a fragment thereof as an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8 , Ab9, AblO, Abll, Abl2, Abl3, Abl4, Abl5,

Abl6, Abl7, Abl8, Abl9, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25,

Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, or

IMPÍ

MEXICAN INSTITUTE OF PROHIEV.Ij

INDUSTRIAL

Ab36 and humanized, human, chimeric or single chain versions thereof that specifically bind IL-6.

Also, the present invention relates to novel methods of preventing or treating cachexia, weakness, fatigue and / or fever in a patient in need using anti-IL-6 antibodies, preferably aglycosylated and / or humanized antibodies that they have an elimination half-life of at least about 25 days.

Description of the related art

Weight loss, fatigue, and muscle weakness are very common symptoms in patients with advanced forms of cancer, and these symptoms may worsen as the cancer progresses. Fatigue, weight loss and muscle weakness can have considerable negative effects on the recovery of patients with advanced forms of cancer, for example by altering lifestyles and relationships and affecting the will or ability of patients to continue anti-cancer treatments. Cancer. Known methods of dealing with fatigue, weight loss, and muscle weakness include regular training and exercise routines, methods to conserve patient energy, and treatments that address anemia-induced fatigue and muscle weakness. However, there remains a need in the art for methods and / or treatments to improve fatigue, weight loss, and

MEXICAN INSTITUTE :,;

OF THE PROPERTY Í> c? s

INDUSTRIAL muscle weakness in cancer patients.

Interleukin-6 (hereinafter IL-6) (also known as interferon-p2; B-cell differentiation factor; B-cell stimulating factor-2; hepatocyte stimulating factor; hybridoma growth factor; and plasmacytoma growth factor) is a multifunctional cytokine involved in numerous biological processes such as regulation of the acute inflammatory response, modulation of specific immune responses including B and T lymphocyte differentiation, bone metabolism, thrombopoiesis, epidermal proliferation, differentiation of menstruation, neuronal cells, neuroprotection, aging, cancer, and the inflammatory reaction that occurs in Alzheimer's disease. See A. Papassotiropoulos, et al, Neurobiology of Aging, 22: 863-871 (2001).

IL-6 is a member of a family of cytokines that promote cellular responses through a receptor complex consisting of at least one subunit of the gpl30 signal transducer glycoprotein and the IL-6 receptor (IL-6R) (also known as gp80). IL-6R may also be present in soluble form (sIL-6R). IL-6 binds to IL-6R, which then dimerizes the gpl30 signal transducer receptor. See Jones, SA, J. Immunology, 175: 3463-3468 (2005).

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MEXICAN INSTITUTE CE THE PROPERTY

INDUSTRIAL

In humans, the gene encoding IL-6 is organized into five exons and four introns and maps to the short arm of chromosome 7 at 7p21. IL-6 RNA translation and post-translational processing results in the formation of a 21-28 kDa protein with 184 amino acids in its mature form. See A. Papassotiropoulos, et al, Neurobiology of

Aging, 22: 863-871 (2001).

As expressed in greater detail herein, IL-6 is believed to play an important role in the development of a multitude of diseases and disorders including but not limited to fatigue, cachexia, autoimmune diseases, diseases of the bone system, cancer, coronary heart disease, obesity, diabetes, asthma, Alzheimer's disease and multiple sclerosis. Due to the perceived involvement of IL-6 in a wide variety of diseases and disorders, there remains a need in the art for useful compositions and methods to prevent or treat illnesses associated with IL-6, as well as monitoring methods to identify patients with illnesses or disorders associated with IL-6. Particularly preferred anti-IL-6 compositions are those that possess minimal or minimized side effects when administered to the patient. Compositions or methods that reduce or inhibit diseases or disorders associated with IL-6 are beneficial to the patient in need.

IL-6 function is not restricted to response

immunological since it acts in hematopoiesis, thrombopoiesis, osteoclast formation, provocation of the acute phase hepatic response that results in the increase of C-reactive protein (CRP) and serum amyloid A protein (SAA). It is known to be a growth factor for epidermal keratinocytes, renal mesangial cells, myeloma cells, and plasmacytoma (Grossman et al., 1989 Prot Nati Acad Sci., 86, (16) 6367-6371; Horii et al., 1989, J Immunol, 143, 12, 39493955; Kawano et al., 1988, Nature 332, 6159, 83-85). IL-6 is produced by a wide variety of cell types including monocytes / macrophages, fibroblasts, epidermal keratinocytes, vascular endothelial cells, renal mesangial cells, glial cells, chondrocytes, T and B lymphocytes, and some tumor cells (Akira et al, 1990, FASEB J., 4, 11, 28602867). Except for tumor cells that constitutively produce IL-6, normal cells do not express

IL-6 unless properly stimulated.

Elevated levels of IL-6 have been observed in various types of cancer, including breast cancer, leukemia, ovarian cancer, prostate cancer, pancreatic cancer, lymphoma, lung cancer, renal cell carcinoma, colorectal cancer, and myeloma. multiple (eg, Chopra et al., 2004, MJAFI 60: 45-49; Songur et al., 2004, Tumori 90: 196-200; Blay et al., 1992,

Cancer Research 52: 3317-3322; Nikiteas et al., 2005, World J.

Gasterenterol. 11: 1639-1643; analyzed in Heikkila et al.,

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

2008, Eur J Cancer, 44: 937-945). As described above, IL-6 is known or suspected to play an important role in promoting the proliferation or survival of at least some types of cancer. Furthermore, some of these studies have shown a correlation between IL-6 levels and the patient's clinical outcome. Taken together, these results suggest the possibility that IL-6 inhibition may be beneficial from a therapeutic point of view.

In fact, clinical studies (analyzed in Trikha et al., 2003,

Clinical Cancer Research 9: 4653-4665) have shown some improvement in the clinical outcome of the patient due to the administration of various anti-IL-6 antibodies, particularly in those cancers in which IL-6 plays a direct role in promoting cancer cell proliferation or survival.

As described above, IL-6 stimulates the acute phase liver response, resulting in increased CRP production and elevated serum CRP levels.

For this reason, C-reactive protein (CRP) has been reported to comprise a surrogate marker for IL6 activity. Therefore, it is possible to detect elevated IL-6 activity by measuring serum CRP. Rather, it is possible to detect effective suppression of IL-6 activity, eg, by administration of a neutralizing anti-IL-6 antibody, by decreasing

IMPi

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INDUSTRIAL

resulting in serum CRP levels.

A recent clinical study showed that administration of rosuvastatin to apparently healthy individuals with elevated CRP (greater than 2.0 mg / 1) reduced their CRP levels by 37% and greatly decreased the incidence of myocardial infarction, stroke, arterial revascularization, hospitalization for unstable angina, or death from cardiovascular causes. Ridker et al.,

N Engl J Med. November 9, 2008 [Electronic publication before printing].

In addition to its direct role in the pathogenesis of some cancers and other diseases, chronically elevated levels of IL-6 appear to adversely affect the well-being and quality of life of patients. For example, elevated IL-6 levels have been reported to be associated with cachexia and fever, and decreased serum albumin. Gauldie et al., 1987, PNAS 84: 7251-7253; Heinric et al., 1990,

265: 621-636; Zamir et al., 1993, Metabolism 42: 204-208; Zamir et al., 1992, Arch Surg, 127: 170-174. Inhibition of IL-6 by a neutralizing antibody has been reported to improve fever and cachexia in cancer patients, although improvement in serum albumin level has not been reported in these patients (Emille et al., 1994, Blood , 84: 2472-2479; Blay et al., 1992, Cancer Research 52: 3317-3322; Bataille et al.,

1995, Blood, 86: 685-691).

IMPIfé

MEXICAN INSTITUTE L) £ INDUSTRIAL PROPERTY

Numerous studies have suggested that CRP is a valuable

Oct; 14 (10): 999-1011, l; 107 (11): 2698-705).

prognostic factor in cancer patients, with elevated CRP levels that predict a poor clinical outcome. See, for example, Hefler et al, Clin Cáncer Res, 2008 Feb 1, -14 (3): 7104; Nagaoka et al, Liver Int, 2007 Oct; 27 (8): 1091-7; Heikkilá et al, J Epidemiol Community Health, 2007 Sep; 61 (9): 824-33,

Review; Hara et al, Anticancer Res, 2007 Jul-Aug; 27 (4C): 3001-4;

Polterauer et al, Gynecol Oncol, 2007 Oct; 107 (1): 114-7, Epub

2007 Jul 6; Tingstedt et al, Scand J Gastroenterol, 2007

Jun; 42 (6): 754-9; Suh et al, Support Care Cancer, 2007

Jun; 15 (6): 613-20, Epub 2007 Jan 18; Gerhardt et al, World J

Gastroenterol, 2006 Sep 14; 12 (34): 5495-500; McArdle et al, Urol Int, 2006; 77 (2): 127-9; Guillem et al, Dis Esophagus,

2005, -18 (3): 146-50; Brown et al, Cancer, 2005 Jan 15; 103 (2): 37782. Decreased serum albumin (hypoalbuminemia) is also associated with increased morbidity and mortality in various serious diseases, including cancers (eg, Vigano et al., Arch Intern Med, 2000 Mar

27; 160 (6): 861-8; Hauser et al., Support Care Cancer, 2006

Seve et al., Cancer, 2006 Dec

The apparent link between hypoalbuminemia and the patient's poor clinical outcome suggests that restoring albumin levels by direct infusion of albumin could promote patient survival, however, albumin infusion has not been

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

improved survival of patients with advanced cancer (Demirkazik et al., Proc Am Soc Clin Oncol 21: 2002 (abstr

2892)) or other groups of seriously ill patients (discussed in Wilkes et al., Ann Intern Med, 2001 Aug

7;135(3):149-64).

The Glasgow Prognostic Score

Glasgow] (GPS) is an inflammation-based prognostic scale that combines albumin (<35 mg / L = 1 point) and CRP (> 10 mg / L = 1 point) (Forrest et al., Br J Cancer , 2004 May 4; 90 (9): 1704-6). Since its introduction in

2004, it has already been shown that the forecast scale of

Glasgow has prognostic value as an indicator of mortality in numerous cancers, including gastro-esophageal cancer, non-small cell lung cancer, colorectal cancer, breast cancer, ovarian cancer, bronchogenic cancer, and metastatic kidney cancer (Forrest et al., Br J Cancer, May 4, 2004; 90 (9): 1704-6; Sharma et al., Clin

Colorectal Cancer, September 2008; 7 (5): 331-7, - Sharma et al., Eur J Cancer, January 2008; 44 (2): 251-6; McMillan et al.,

Nutr Cancer, 2001; 41 (1-2): 64-9; McMillan, Proc Nutr Soc, August 2008; 67 (3): 257-62, - Ramsey et al., Cancer, January 15,

2007,-109(2) :205-12) .

US Patent Application Publication No.

20080081041 (related to cancer treatment using an anti-IL-6 antibody) describes that because IL-6 is

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MEXICAN INSTITUTE OF THE

1N üUSTkiAL

associated with disease activity and since CRP is a surrogate marker for IL-6 activity, sustained suppression of CRP by neutralizing IL-6 through its anti-IL-6 antibody ( CNTO 328, Zaki et al.r Int J

Cancer, 2004 Sep 10; 111 (4): 592-5) may be assumed necessary to achieve biological activity. The same patent application indicates that the relationship between IL-6 and CRP in patients with benign and malignant prostate disease was previously examined by McArdle (McArdle et al. 2004 Br J Cancer

91 (10): 1755-1757). McArdle reportedly found no significant difference between IL-6 and

CRP in patients with benign disease compared to patients with prostate cancer, in cancer patients there was a considerable increase in both the concentration of IL-6 and CRP with an increasing degree of tumor. The mean serum CRP value for the 8 6 subjects with prostate cancer was 1.8 mg / L. Based on this, the inventors of the present patent application postulate a proposed dose and schedule where 6 mg / kg of an anti-IL-6 antibody (CNTO 328) is administered every 2 weeks and state that this is likely to achieve sustained suppression of CRP in subjects with metastatic HRPC.

IL-6 signaling is mediated by the Jak-Tyk family of cytoplasmic tyrosine kinases, including JAK1, JAK2, and JAK3 (discussed in Murray J Immunol. March 1,

Mexican institute OF PROPERTY

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2007; 178 (5): 2623-9). Sivash et al. report the abrogation of IL-6 mediated JAK signaling by prostaglandin cyclopentenone 15d-PGJ2 in oral squamous cell carcinoma cells. British Journal of Cancer (2004) 91, 1074-1080. These results suggest that inhibitors of JAK1, JAK2, or JAK3 could be used as IL-6 antagonists.

Ulanova et al. report that inhibition of the non-receptor protein tyrosine kinase Syk (using siRNA) decreased IL-6 production by epithelial cells. Am J Physiol Lung Cell Mol Physiol March 2005; 288 (3): L497-507. These results suggest that a Syk inhibitor could be used as an IL-6 antagonist.

Kedar et al. report that thalidomide treatment significantly reduces serum levels of CRP and IL-6 to normal or near normal levels in a significant fraction of patients with renal cell carcinoma. Int J

Cancer. June 10, 2004; 110 (2): 260-5. These results suggest that thalidomide and possibly derivatives thereof, such as lenalidomide, could be useful IL-6 antagonists.

Also, another published patent application, USA

20070292420 teaches a Phase I dose escalation study using an anti-IL-6 antibody (cCLB-8) to treat patients who do not respond to treatment with advanced stage multiple myeloma (N = 12) and indicates that this study demonstrates that some

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY patients presented a stabilization of their disease. The application also informs that after the interruption of the treatment there was an acceleration in the increase of the levels of protein M, which suggests a rebound of the disease after the cessation of the treatment. The anti-IL-6 antibody cCLB-8 inhibited freely circulating IL-6.

The application also indicates that this antibody assay did not result in toxicity (except transient thrombocytopenia in two previously intensively treated patients) or allergic reactions and decreased C-reactive protein (CRP) below detection level in all patients. . Their antibody (cCLB-8 antibody) allegedly possessed a circulating half-life of 17.8 days and no immunological response to human anti-chimeric antibody (HACA) was observed (van Zaanen et al. 1998). They claim that the administration of CNTO 328 did not cause changes in blood pressure, heart rate, temperature, hemoglobin, liver functions, and kidney functions. Except for transient thrombocytopenia in two previously intensively treated patients, no toxicity or allergic reactions were reportedly observed, and no immunological response to human anti-chimeric antibody (HACA) was observed. Three patients in their study allegedly developed infection-related complications during treatment, however, the inventors concluded

IΜ ΡI

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL _ that a possible relationship with the anti-IL-6 antibody cCLB-8 was unlikely because infectious complications are supposedly common in end-stage multiple myeloma and are a leading cause of death. Based on their results, they concluded that this anti-IL-6 cCLB-8 antibody was safe in patients with multiple myeloma.

As previously stated, elevated IL-6 was implicated in the pathogenesis of cachexia, weakness, fatigue, and fever. The diseases and disorders associated with fatigue include, but are not limited to, general fatigue, exercise-induced fatigue, cancer-related fatigue, inflammatory disease-related fatigue, and chronic fatigue syndrome. See, for example, Esper DH, et al, The cancer cachexia syndrome: a review of metabolic and clinical manifestations, Nutr Clin Pract., August 2005, -20 (4): 369-76; Vgontzas AN, et al, IL-6 and its circadian secretion in humans,

Neuroimmunomodulation, 2005; 12 (3).-131-40; Robson-Ansley, PJ, et al, Acute interleukin-6 administration impairs athletic performance in healthy, trained male runners, Can J Appl

Physiol., August 2004; 29 (4): 411-8; Shephard RJ., Cytokine responses to physical activity, with particular reference to

IL-6: sources, actions, and clinical implications, Crit Rev Immunol., 2002; 22 (3): 165-82, · Arnold, MC, et al, üsing an interleukin-6 challenge to evalúate neuropsychological performance in chronic fatigue syndrome , Psychol Med., August

2002; 32 (6): 1075-89; Kurzrock R., The role of cytokines in cancer-related fatigue, Cancer, September 15, 2001/92 (6 Suppl): 1684-8; Nishimoto N, et al, Improvement in Castleman's disease by humanized anti-interleukin-6 receptor antibody therapy, Blood, January 1, 2000; 95 (1): 56-61; Vgontzas AN, et ai, Circadian interleukin-6 secretion and quantity and depth of sleep, J Clin Endocrinol Metab., August 1999; 84 (8): 26037 and Spath-Schwalbe E, et ai, Acute effects of recombinant human interleukin 6 on endocrine and central nervous sleep functions in healthy men, J Clin Endocrinol Metab., May 1998;

83 (5): 1573-9, the disclosures of which are incorporated herein by reference in their entirety.

The diseases and disorders associated with cachexia include, but are not limited to, cancerous cachexia, cardiac cachexia, respiratory cachexia, renal cachexia, and age-related cachexia. See, for example, Barton, BE., Interleukin-6 and new strategies for the treatment of cancer, hyperproliferative diseases and paraneoplastic syndromes, Expert Opin Ther Targets, August 2005; 9 (4): 73752; Zaki MH, et al, CNTO 328, a monoclonal antibody to IL-6, inhibíts human tumor-induced cachexia in nude mice, Int J

Cancer, September 10, 2004; 111 (4): 592-5; Trikha M, et al, Targeted anti-interleukin-6 monoclonal antibody therapy for cancer: a review of the rationale and clinical evidence, Clin

Cancer Res., October 15, 2003/9 (13): 4653-65; Lelli G, et

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MEXICAN INSTITUTE ~ 'Z,', Ί · <

FROM INDUSTRIAL PROPERTY al, Treatment of the cancer anorexia-cachexia syndrome: a critical reappraisal, J Chemother., June 2003; 15 (3): 220-5; Argües JM, et al, Cytokines in the pathogenesis of cancer cachexia, Curr Opin Clin Nutr Metab Care, July 2003;

6 (4): 401-6, - Barton BE., IL-6-like cytokines and cancer cachexia: consequences of chronic inflammation, Immunol Res.,

2001; 23 (1) .- 41-58; Yamashita JI, et al, Medroxyprogesterone acétate and cancer cachexia: interleukin-6 involvement, Breast

Cancer, 2000; 7 (2) .Ί30-5; Yeh SS, et al, Geriatric cachexia: the role of cytokines, Am J Clin Nutr., August 1999; 70 (2): 18397; Strassmann G, et al, Inhibition of experimental cancer cachexia by anti-cytokine and anti-cytokine-receptor therapy, Cytokines Mol Ther., June 1995; l (2): 107-13; Fujita J, et al, Anti-interleukin-6 receptor antibody prevents muscle atrophy in colon-26 adenocarcinoma-bearing mice with modulation of lysosomal and ATP-ubiquitin-dependent proteolytic pathways, Int J Cáncer, November 27, 1996; 68 (5): 637-43; Tsujinaka T, et al, Interleukin 6 receptor antibody inhibits muscle atrophy and modulates proteolytic systems in interleukin 6 transgenic mice, J Clin Invest., January 1, 1996; 97 (1): ·, 244-9; Emilie

D, et al, Administration of an anti-interleukin-6 monoclonal antibody to patients with acquired immunodeficiency syndrome and lymphoma: effect on lymphoma growth and on B clinical

Symptoms, Blood, October 15, 1994; 84 (8): 2472-9, - and

Strassmann G, et al, Evidence for the involvement of

INSTITUTO MEXICANO D2 LA PROPIEDAD INDUSTRIAL interleukin 6 in experimental cancer cachexia, J Clin Invest., May 1992; 89 (5): 1681-4; the descriptions of which are incorporated herein by reference in their entirety.

Another disease related to cachexia is decline syndrome, also known as growth retardation, where a child has a lower than expected rate of weight gain. Decline syndrome is typically defined as a weight below the third percentile or a decrease in the percentile range of 2 main growth parameters in a limited period. Decline syndrome occurs as a result of heterogeneous medical and psychosocial causes, and sometimes the cause eludes the diagnosis. A recent study (with a total of 34 patients) indicated statistically significant growth in IL-6 levels in patients diagnosed with decline syndrome. Shaoul et al. J Pediatr Gastroenterol

Nutr., October 2003; 37 (4); 487-91.

BRIEF SUMMARY OF THE INVENTION

The present invention is an extension of applicants' prior inventions directed at specific antibodies, humanized or chimeric or single-chain antibodies and fragments thereof that possess binding specificity for IL-6, in particular antibodies that possess

MEXICAN INSTITUTE of property

INDUSTRIAL

Specific epitope specificity and / or functional properties and novel therapies using these and other anti-IL-6 antibodies. An embodiment of the invention comprises specific humanized antibodies and fragments thereof capable of binding IL-6 and / or the IL-6 / IL-6R complex. These antibodies can bind soluble IL-6 or IL-6 expressed on the cell surface. Also, these antibodies can inhibit the formation or biological effects of one or more IL-6s, IL-6 / IL-6R complexes, IL-6 / IL-6R / gpl30 complexes and / or IL6 / IL-6R / multimers. gpl30. The present invention relates to novel therapies and therapeutic protocols using anti-IL-6 antibodies, preferably those described herein. In particular, the present invention relates to methods of preventing or treating cachexia, weakness, fatigue, and / or fever in a patient in need, eg, a patient displaying elevated levels of CRP, including administering to the patient an anti-IL-6 antibody or antibody fragment, whereby cachexia, weakness, fatigue and / or fever of the patient is prevented or ameliorated, or restored to a normal state.

In a preferred embodiment this is done by administering the antibodies described herein, which comprise the V polypeptide sequences.H, VL and CDRs described herein, or humanized or chimeric or single chain versions thereof containing one or more of the

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CDRs of the exemplified anti-IL-β antibody sequences and the polynucleotides encoding them. Preferably these antibodies will be aglycosylated. In more specific embodiments of the invention these antibodies will block gpl30 activation and / or possess binding affinities (Kds) less than 50 picomolar and / or K valuesoff less than or equal to 10 ~4 S_1.

In another embodiment of the invention these antibodies and humanized versions will come from rabbit immune cells (B lymphocytes) and can be selected based on their homology (sequence identity) with human germline sequences. These antibodies may require minimal sequence modifications, or may not require modifications, thereby facilitating the retention of functional properties after humanization. In exemplary embodiments these humanized antibodies will comprise human flanking regions that are highly homologous (possess high levels of sequence identity) to those of a parent antibody (eg rabbit) as described below.

In another embodiment of the invention the antibodies in question can be selected according to their activity in functional assays such as IL-6 driven T1165 proliferation assays, IL-6 simulated HepG2 cell haptoglobin production assays, and the like. A further embodiment of the invention is directed to anti-IL-6 antibody fragments comprising V polypeptides.H, VL and

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CDRs, eg, that come from rabbit immune cells and the polynucleotides that encode them, as well as the use of these antibody fragments and the polynucleotides that encode them in the creation of novel antibodies and polypeptide compositions capable of recognizing complex IL-6 and / or IL-6 / IL-6R or IL-6 / IL-6R / gpl30 complexes and / or multimers thereof.

The invention also contemplates administration of anti-IL-6 antibody conjugates and humanized, chimeric, or single chain versions thereof and other binding fragments of the same conjugates to one or more functional or detectable moieties. The invention also contemplates methods for making such humanized anti-IL-6 antibodies or anti-IL-6 / IL-6R complex antibodies and binding fragments thereof. In one embodiment, the junction fragments include but are not limited to Fab, Fab ', F (ab') fragments2, Fv and scFv.

The embodiments of the invention relate to the use of anti-IL-6 antibodies for the diagnosis, evaluation and treatment of diseases and disorders associated with IL-6 or the aberrant expression thereof. The invention also contemplates the use of anti-IL-6 antibody fragments for the diagnosis, evaluation and treatment of diseases and disorders associated with IL-6 or the aberrant expression thereof. The preferred uses of the antibodies in question,

I especially humanized, chimeric and single chain antibodies are the treatment and prevention of fatigue associated with cancer and / or cachexia and rheumatoid arthritis.

Other embodiments of the invention relate to the production of anti-IL-6 antibodies in recombinant host cells, preferably diploid yeast such as

Diploid pichia and other yeast strains.

Another embodiment of the invention relates to methods for improving the survivability and quality of life of a patient diagnosed with cancer, comprising administering to the patient an anti-IL-6 antibody or antibody fragment, whereby the patient's serum C-reactive protein (CRP) level is stabilized and preferably reduced, and the patient is monitored to assess the reduction in the patient's serum CRP level, where the anti-IL-6 antibody or antibody fragment can specifically bind to the same linear or conformational epitope (s) and / or compete to bind to the same epitope (s) linear or conformational epitope (s) in an intact human IL-6 polypeptide or fragment thereof as an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9,

<td>Ab30,</td><td>Ab31,</td><td>Ab32,</td><td>Ab33,</td><td>Ab34, Ab35, or Ab36 and</td><td colspan="2">antibodies</td>

chimeric, humanized, single-chain and fragments of

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themselves (containing one or more CDRs of the antibodies mentioned above) that specifically bind to IL-6, which are preferably aglycosylated.

Another embodiment of the invention relates to methods of improving the muscle strength of a patient diagnosed with cancer, which comprises administering to the patient an anti-IL-6 antibody or antibody fragment, thereby improving the patient's muscle strength. , and monitor the patient to assess muscle strength, where the anti-IL-6 antibody or antibody fragment can specifically bind to the same linear or conformational epitope (s) and / or compete to bind to the same epitope (s) linear or conformational epitope (s) in an intact human IL-6 polypeptide or fragment thereof as an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9,

<td>Ab30, Ab31,</td><td>Ab32,</td><td>Ab33</td><td>, Ab34,</td><td>Ab35, or Ab36 and antibodies</td>

<td>chimeric,</td><td colspan="2">humanized,</td><td colspan="2">single chain and fragments of</td>

<td>themselves (that</td><td colspan="2">they contain</td><td>one or</td><td>more CDR from the antibodies</td>

mentioned above) that specifically bind to IL6, which are preferably aglycosylated. In such methods preferably the muscle strength of the patients improves by at least about 15% within approximately 4 weeks from the administration of the antibody or fragment of

IΜ ΡI

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OF THE PROPERTY

INDUSTRIAL anti-IL-6 antibody, as measured by the Hand Grip test

Strength and most preferably muscle strength of patients improves by at least about 20% within approximately 4 weeks from administration of the anti-IL-6 antibody or antibody fragment, as measured by the test. grip force.

Another embodiment of the invention relates to methods of increasing serum albumin in a patient in need, comprising administering to the patient an anti-IL-6 antibody or antibody fragment, thereby increasing serum albumin from the patient, and monitor the patient to assess the serum albumin level, where the anti-IL-6 antibody or antibody fragment can specifically bind to the same linear or conformational epitope (s) and / or compete to bind to the same epitope (s) linear or conformational epitope (s) in an intact human IL-6 polypeptide or fragment thereof as an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9,

<td>Ab30,</td><td>Ab31,</td><td>Ab32, Ab33</td><td>, Ab34,</td><td>Ab35, or Ab36 and</td><td>antibodies</td>

<td colspan="2">chimeric,</td><td>humanized,</td><td colspan="3">single chain and fragments of</td>

<td>themselves</td><td>(than</td><td>they contain</td><td>one or</td><td>more CDR than</td><td>antibodies</td>

mentioned above) that specifically bind to IL-6, which are preferably aglycosylated. Preferably these

methods are carried out under conditions whereby the patient's survivability is improved and / or under conditions where the serum albumin level increases by about 5 g / L, within approximately 6 weeks from the administration of the antibody or antiIL-6 antibody fragment. These patients will include, without limitation to them, those diagnosed with rheumatoid arthritis, cancer, advanced cancer, liver disease, kidney disease, inflammatory bowel disease, celiac disease, trauma, burns, other diseases associated with decreased albumin in serum or any combination thereof.

An embodiment of the invention relates to methods of preventing or treating cachexia, weakness, fatigue and / or fever in a patient diagnosed with an IL-6 associated disorder, comprising administering to the patient an antibody or antibody fragment. anti-IL-6, thereby preventing or improving cachexia, weakness, fatigue and / or fever of the patient, and monitoring the patient to assess cachexia, weakness, fatigue and / or fever, where the anti-IL-6 antibody or antibody fragment can specifically bind to the same linear or conformational epitope (s) and / or compete to bind to the same (s) linear or conformational epitope (s) in an intact human IL-6 polypeptide or fragment thereof as an antibody

Ab7, Ab8,

ΡΙ

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL anti-IL-6 comprising Abl, Ab2, Ab3, Ab4, Ab5, Ab6,

<td colspan="2">Ab29, Ab30,</td><td>Ab31,</td><td>Ab32,</td><td>Ab33, Ab34, Ab35, or</td><td colspan="2">Ab36 and antibodies</td>

chimeric, humanized, single chain and fragments thereof (containing one or more CDRs of the antibodies mentioned above) that specifically bind to IL-6, which are preferably aglycosylated. As discussed below, in a preferred embodiment the anti-IL-6 antibody will comprise a humanized antibody containing the

CDR of Abl and more preferably will comprise the variable heavy and light chain in SEQ ID NO: 657 and SEQ ID NO: 709 respectively and the constant regions in SEQ ID NO: 588 and 586 respectively or variants thereof where one or more Amino acids are modified by substitution or deletion without substantially altering the binding affinity for IL-6.

In a preferred embodiment the humanized anti-IL-6 antibody will comprise the variable heavy chain and variable light chain sequences respectively contained in the

SEQ ID NO: 657 and SEQ ID NO: 709, and preferably further comprising the heavy and light chain constant regions respectively contained in SEQ ID NO: 588 and SEQ ID NO: 586, and variants thereof comprising one or plus amino acid substitutions or deletions that do not substantially affect IL-6 binding and / or the desired effector function. The present modality

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL too

contemplates polynucleotides that comprise, or alternatively consist of, one or more nucleic acids encoding the variable heavy chain (SEQ ID NO: 700) and variable light chain (SEQ ID NO: 723) sequences and the region sequences heavy chain constant (SEQ ID NO: 58 9) and light chain constant region (SEQ ID NO: 587). This embodiment further contemplates nucleic acids encoding variants comprising one or more amino acid substitutions or deletions to the variable heavy and light chain sequences respectively contained in SEQ ID NO: 657 and SEQ ID NO: 709 and the heavy chain constant regions and light chain respectively contained in SEQ ID NO: 588 and SEQ ID NO: 586, which do not substantially affect IL-6 binding and / or desired effector function.

In one embodiment of the invention, the anti-IL-6 antibody can bind to the same linear or conformational epitope (s) and / or compete to bind to the same epitope (s) ) linear or conformational epitope (s) in an intact human IL-6 polypeptide or fragment thereof as Abl.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can specifically bind to the same linear or conformational epitope (s) in an intact IL-6 polypeptide human or fragment thereof as an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4,

ΙΜΡΪ

INSTITUTE Μ.:· '? '.'- /. VO DE LA f? C:: INuUSTiilAL

Ab5, Ab6, Ab7, Ab8, Ab9, AblO, Abll, Abl2, Abl3, Abl4, Abl5,

Abl6, Abl7, Abl8, Abl9, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25,

Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, or Ab36 and single chain humanized chimeric antibodies and fragments thereof (containing one or more CDRs of the aforementioned antibodies) that specifically bind to the

IL-6, which are preferably aglycosylated.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can specifically bind to the same linear or conformational epitope (s) in an intact IL-6 polypeptide human or a fragment thereof such as Abl or a humanized or chimeric antibody comprising all or most of the same CDRs as Abl that specifically bind IL-6.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can specifically bind to the same linear or conformational epitopes on an intact IL-6 polypeptide or antibody fragment thereof that is (are) specifically bound by Abl and where said epitope (s), when established by epitopic mapping using overlapping linear peptide fragments spanning the entire length of native human IL-6 polypeptide, include one or more residues comprised of IL-6 fragments that are selected from those that respectively comprise amino acid residues 37-51, amino acid residues 70-84, residues

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INSTITUTO .MEXICANO LA VKCFÍEÜAD INDUSTRIAL

of amino acids 169-183, amino acid residues 31-45 and / or amino acid residues 58-72.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment may comprise at least 2 Complementarity Determining Regions (CDRs) in each variable light and variable heavy region that are identical to those contained in an anti-IL antibody. -6 comprising

<td>Ab33,</td><td>Ab34,</td><td>Ab35,</td><td colspan="2">or Ab36 or a combination</td><td>of</td><td>CDR of</td><td>one</td>

several of said antibodies.

In one embodiment of the invention, all CDRs in the anti-IL-6 antibody or antibody fragment can be identical to CDRs contained in an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4, Ab5, Ab6 , Ab7, Ab8, Ab9, AblO, Abll, Abl2, Abl3, Abl4, Abl5, Abl6, Abl7, Abl8, Abl9, Ab20,

Ab21, Ab22, Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30,

Ab31, Ab32, Ab33, Ab34, Ab35, or Ab36 and humanized, single-chain chimeric antibodies and fragments thereof (which

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MEXICAN INSTITUTE D £ u \ INDUSTRIAL PROPERTY

they contain one or more CDRs of the antenomono-monion ^ -— antibodies that specifically bind to IL-6, which are preferably aglycosylated.

Another embodiment of the invention relates to Abl, including humanized and rabbit forms thereof, as well as heavy chains, light chains, fragments, variants, and CDRs thereof. In the human clinical trials presented in the Examples, a humanized form of Abl was administered.

In one embodiment of the invention, all CDRs in the anti-IL-6 antibody or antibody fragment can be identical to the CDRs contained in Abl.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can be aglycosylated.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment may contain an Fe region that has been modified to alter effector, half-life, proteolysis and / or glycosylation function. Preferably the Fe region is modified to eliminate glycosylation.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can be a human, humanized, single chain or chimeric antibody.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can be a humanized antibody that comes from a rabbit anti-IL-6 antibody

ΙΜΡ1

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY (parent).

In one embodiment of the invention, the flanking regions (FR) in the variable light region and variable heavy regions of said anti-IL-6 antibody or antibody fragment respectively can be unmodified human FR or have been modified by substitution of at most 2 or 3 human FR residues in the variable light or heavy chain region with the corresponding residues of

FR of the rabbit parent antibody, and human FRs can come from variable heavy and light chain antibody sequences that have been selected from a library of human germline antibody sequences based on their high level of homology with the corresponding heavy chain regions or slight rabbit variables relative to other human germline antibody sequences contained in the library. As described in detail below, in a preferred embodiment the antibody will comprise human FRs that are selected based on their high level of homology (degree of sequence identity) to those of the main antibody that is humanized.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can be administered to the patient at a frequency of at most once per period of approximately four weeks, approximately eight weeks, approximately twelve weeks, approximately sixteen weeks, approximately approximately twenty four weeks.

Prevent

MEXICAN INSTITUTE Y '”- X-'1

OF PROPERTY Λ J, INDUSTRIAL twenty weeks or

In one embodiment of the invention, improvement in the patient's cachexia, weakness, fatigue and / or fever can be maintained for a full period between two consecutive administrations of the anti-IL-6 antibody.

In one embodiment of the invention, the patient may have been diagnosed with selected acanthoma cancer, acinic cell carcinoma, acoustic neuroma, acral lentiginous melanoma, acrospiroma, acute eosinophilic leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monocytic leukemia, leukemia Acute myeloblastic with maturation,

-w dendritic cell acute myeloid leukemia, acute myeloid leukemia, acute promyelocytic leukemia, adamantinoma, adenocarcinoma, adenoid cystic carcinoma, adenoma, adenomatoid odontogenic tumor, adult T-cell leukemia, aggressive NK-cell leukemia, cancers related to AIDS, AIDS-related lymphoma, alveolar soft tissue sarcoma, ameloblastic fibroma, anal cancer, anaplastic large cell lymphoma, Anaplastic thyroid cancer, T-cell angioimmunoblastic lymphoma, angiomyolipoma, angiosarcoma, appendix cancer, astrocytoma, atypical rhabdoid teratoid tumor, basal cell carcinoma, B-cell leukemia, B-cell lymphoma, duct carcinoma by Bellini,

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TO MEXICAN INSTITUTE OF THE PROi'lEOAO v ------ INDUSTRIAL biliary tract cancer, bladder cancer, blastoma, bone cancer, bone tumor, brain stem glioma, brain tumor, breast cancer, Brenner's tumor, bronchial tumor, bronchioloalveolar carcinoma, tumor of Brown lymphoma

Burkitt, cancer of unknown primary site, carcinoid tumor, carcinoma, carcinoma in situ, carcinoma of the penis, carcinoma of unknown primary site, carcinosarcoma, Castleman's disease, embryonal tumor of the central nervous system, cerebellar astrocytoma, cerebral astrocytoma, cervical cancer, cholangiocarcinoma , chondroma, chondrosarcoma, chordoma, choriocarcinoma, choroid plexus papilloma, chronic lymphocytic leukemia, chronic monocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorder, chronic neutrophilic leukemia, clear cell tumor, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, Degos disease, dermatofibrosarcoma protuberans, dermoid cyst, desmoplastic small-cell tumor, diffuse B-cell lymphoma large, dysmbrioplastic neuroepithelial tumor, embryonal carcinoma, endodermal sinus tumor, endometrial cancer, endometrial uterine cancer, endometrioid tumor, T-cell lymphoma associated with enteropathy, ependymoblastoma, ependymoma, epithelioid, erythroleukemia, cancer of esthesioneuroblastoma, Ewing family tumor, Ewing family sarcoma, Ewing sarcoma, extracranial tumor of esophageal sarcoma, or

I

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

.VI Germ Cells, Extragonadal Germ Cell Tumor, Extrahepatic Bile Duct Cancer, Extramammary Paget's Disease, Fallopian Tube Cancer, Fetus in fetu, Fibroma, Fibrosarcoma, Follicular Lymphoma, Follicular Thyroid Cancer, Gallbladder Cancer, gallbladder, ganglioglioma, ganglioneuroma, gastric cancer, gastric lymphoma, gastrointestinal cancer, gastrointestinal carcinoid tumor, carcinoid tumor, gastrointestinal stromal tumor, gastrointestinal stromal tumor, germ cell tumor, germinoma, gestational choriocarcinoma, gestational trophoblastic tumor, giant cell bone tumor, glioblastoma multiforme, glioma, gliomatosis cerebri, glomus tumor, glucagonoma, gonadoblastoma, granule cell tumor, villous cell leukemia, hairy cell leukemia, head and neck cancer, head and neck cancer, heart cancer, hemangioblastoma, hemangiopericytoma, hemangiosarcoma, hematologic malignancy, hepatocellular carcinoma, hepatosplenic T-cell lymphoma, hereditary ovarian cancer syndrome, Hodgkin lymphoma, Hodgkin lymphoma, hypopharyngeal cancer, hypothalamic glioma, inflammatory breast cancer, islet cell carcinoma, tumor of islet cells, juvenile myelomonocytic leukemia, Kaposi sarcoma, Kaposi sarcoma, kidney cancer, Klatskin tumor, Krukenberg tumor, laryngeal cancer, laryngeal cancer, melanoma

MEXICAN INSTITUTE \ 3

FROM LAFROPUVAD VW

IN DUSTRIAL Malignant lentigo, leukemia, leukemia, cancer of the lip and oral cavity, liposarcoma, lung cancer, luteoma, lymphangioma, lymphangiosarcoma, lymphoepithelioma, lymphoid leukemia, lymphoma, macroglobulinemia, fibrous malignant histiocytoma, fibrous malignant histiocytoma , malignant glioma, malignant mesothelioma, malignant peripheral nerve sheath tumor, malignant rhabdoid tumor, malignant newt tumor, MALT lymphoma, mantle cell lymphoma, mast cell leukemia, mediastinal germ cell tumor, mediastinal tumor, medullary thyroid cancer, medulloblastoma, medulloblastoma, medulloepithelioma, melanoma, melanoma, meningioma, Merkel cell carcinoma, mesothelioma, mesothelioma, metastatic squamous neck cancer with occult primary tumor, metastatic urothelial carcinoma, mixed Müllerian tumor, monocytic leukemia, mouth cancer, mucinous tumor, multiple endocrine neoplasia syndrome, multiple myeloma, multiple myeloma, mycosis fungoides, mycosis fungoides, myelodysplastic disease, myelodysplastic syndromes, myeloid leukemia, myeloid sarcoma, myeloproliferative disease, myxoma, cancer of the nasal cavity, nasopharyngeal cancer, nasopharyngeal carcinoma, neoplasm, neurinoma, neuroblastoma, Nodular melanoma, non-Hodgkin lymphoma, non-Hodgkin lymphoma, non-melanoma skin cancer, non-small cell lung cancer, ocular oncology, oligoastrocytoma,

IMPI

MEXICAN INSTITUTE D £ LA PRO ?! F.'Ali INDUSTRIAL oligodendroglioma, oncocytoma, meningioma of the optic nerve sheath, oral cancer, oral cancer, oropharyngeal cancer, osteosarcoma, osteosarcoma, ovarian cancer, ovarian cancer, ovarian epithelial cancer, germ cell tumor of the ovary , low malignant potential ovarian tumor, Paget disease of the breast, Pancoast tumor, pancreatic cancer, pancreatic cancer, papillary thyroid cancer, papillomatosis, paraganglioma, paranasal sinus cancer, parathyroid cancer, penile cancer, perivascular epithelioid cell tumor, pharyngeal cancer, pheochromocytoma, pineal parenchymal tumor of intermediate differentiation, pineoblastoma, pituicytoma, pituitary adenoma, pituitary tumor, plasma cell neoplasm, pleuropulmonary blastoma , precursor lymphoblastic T-cell lymphoma, primary lymphoma of the central nervous system, primary effusion lymphoma, primary hepatocellular cancer, primary liver cancer, primary peritoneal cancer, primitive neuroectodermal tumor, prostate cancer, peritoneal pseudomyxoma, rectal cancer, renal cell carcinoma, NUT gene related respiratory tract carcinoma in croomosoma 15, retinoblastoma, rhabdomyoma, rhabdomyosarcoma, Richter transformation , sacrococcygeal teratoma, salivary gland cancer, sarcoma, Schwannomatosis, sebaceous gland carcinoma, secondary neoplasm, seminoma, serous tumor, Sertoli-Leydig cell tumor, sex cord stromal tumor,

IMPI

M.LXICai-IO INSTITUTE OF ΙΛ INDUSTRIAL PROPERTY Sézary syndrome, signet ring cell carcinoma,

INDUSTRIAL skin cancer, round and blue small cell tumor, small cell carcinoma, small cell lung cancer, small cell lymphoma, small bowel cancer, soft tissue sarcoma, somatostatinoma, chimney sweep wart, spinal cord tumor , spinal tumor, splenic marginal zone lymphoma, squamous cell carcinoma, stomach cancer, disseminating superficial melanoma, supratentorial primitive neuroectodermal tumor, superficial epithelial ovarian tumor, synovial sarcoma, acute T-cell lymphoblastic leukemia, T-cell granular large lymphocytic leukemia, T-cell leukemia, T-cell lymphoma, teratoma, terminal lymphatic cancer, testicular cancer, tecoma , throat cancer, thymic carcinoma, thymoma, thyroid cancer, transitional cell cancer of the renal pelvis and ureter, transitional cell carcinoma, urachal cancer, urethral cancer, urogenital neoplasm, uterine sarcoma, uveal melanoma, vaginal cancer, Verner Morrison syndrome, warty carcinoma, visual glioma, vulvar cancer, Waldenstrom's macroglobulinemia, Warthin's tumor, Wilms' tumor, or any combination thereof.

In one embodiment of the invention, the patient may have been diagnosed with cancer selected from colorectal cancer, non-small cell lung cancer, cholangiocarcinoma, mesothelioma, or renal cell carcinoma itself.

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY Cas disease any combination 'efe ”lós

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment may comprise a VH polypeptide sequence comprising: SEQ ID NO: 3, 18,

<td> 688,</td><td> 691, 692,</td><td> 704,</td><td colspan="3">or 708 or the VH sequences contained in</td>

<td>the</td><td>antibodies</td><td colspan="2">represented in Figures 34-37</td><td>; and</td><td>can</td>

further comprising a V polypeptide sequenceL than

<td colspan="2">understands:</td><td colspan="3">SEQ ID NO: 2,</td><td> 20, 21,</td><td> 37,</td><td> 53,</td><td> 69, 85,</td><td> 101,</td><td> 119,</td><td> , 122,</td>

<td> 683,</td><td> 687,</td><td> 693,</td><td> 699,</td><td> 702</td><td> , 706,</td><td colspan="2">or 709 or</td><td colspan="3">the sequences</td><td>from VH</td>

contained in the antibodies depicted in Figures 3437 or a variant thereof where one or more of the flanking residues (FR residues) in said V polypeptideH 0 VL it may have been replaced with another amino acid residue resulting in an anti-IL-6 antibody or antibody fragment that specifically binds human IL-6. Preferably the variable heavy and light sequences comprise those in

INSTITUTO MEXICANO DC THE PROPERTY

INDUSTRIAL

SEQ ID NO: 657 and 709. _

In an embodiment of the invention, one or more of said FR residues can be replaced by an amino acid present at the corresponding site in a parental rabbit anti-IL-6 antibody from where the complementarity determining regions (CDR) contained in said VH or VL polypeptides and by a consecutive amino acid substitution.

<td>In</td><td>a</td><td>modality</td><td>of the invention,</td><td>said antibody</td><td>or</td>

<td>fragment</td><td>of</td><td>antibody</td><td>anti-IL-6 can be</td><td>humanized.</td><td></td>

<td>In</td><td>a</td><td>modality</td><td>of the invention,</td><td>said antibody</td><td>or</td>

<td>fragment</td><td>of</td><td>antibody</td><td>anti-IL-6 can be</td><td>chimerical.</td><td></td>

<td>In</td><td>a</td><td>modality</td><td>of the invention,</td><td>said antibody</td><td> 0</td>

<td>fragment</td><td>of</td><td>antibody</td><td colspan="3">anti-IL-6 may further comprise a</td>

Human Fe, eg, a Fe region comprising the variable heavy and light chain constant regions contained in the

SEQ ID NO: 704 and 702.

In an embodiment of the invention, said human Fe can come from IgGl, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8,

IgG9, IgGlO, IgGll, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17,

IgG18 or IgG19.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment may comprise a polypeptide having at least 90% sequence homology to one or more of the polypeptide sequences of SEQ ID

NO: 3, 18, 19, 22, 38, 54, 70, 86, 102, 117, 118, 123, 139,

<td rowspan="3"> 155,</td><td rowspan="3"> 171,</td><td rowspan="3"> 187,</td><td rowspan="3"> 203,</td><td rowspan="3"> 219,</td><td rowspan="3"> 235,</td><td rowspan="3"> 251,</td><td rowspan="3"> 267,</td><td colspan="2" rowspan="2">Mr. Vacano Institute of Industrial Property</td><td colspan="2"></td>

<td colspan="2">sequences</td><td colspan="2">VH and VL</td><td colspan="4">represented in</td><td colspan="2">Figures 34</td><td> -37.</td><td></td>

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can have an elimination half-life of at least about 22 days, at least about 25 days, or at least about 30 days.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment can be co-administered with a chemotherapeutic agent.

In one embodiment of the invention, the chemotherapeutic agent can be selected from VEGF antagonists, EGFR antagonists, platins, taxols, irinotecan, 520 fluorouracil, gemcitabine, leucovorin, spheroids, ——— rufophosp amide, melphalan, vinca alkaloids ( for example, viriblastin, vinC ^^ ípfiSSysa ^ a & ndesine and vinorelbine), mustines, tyrosine kinase inhibitors, radiotherapy, sex hormone inhibitors, selective androgen receptor modulators, selective estrogen receptor modulators,

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PDGF antagonists, TNF antagonists, IL1 antagonists, interleukins (eg IL-12 or IL-2), IL-12R antagonists, toxin-conjugated monoclonal antibodies, specific monoclonal antibodies against tumor antigens, Erbitux ™, Avastin ™ , Pertuzumab, anti-CD20 antibodies, Rituxan®, ocrelizumab, ofatumumab, DXL625, Herceptin® or any combination thereof.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment that can be directly or indirectly linked to a detectable label or therapeutic agent.

In one embodiment of the invention, the anti-IL-6 antibody or antibody fragment may be Abl or humanized, chimeric, single chain or fragment thereof, comprising all or most of the Abl CDRs.

In an embodiment of the invention, the disease or condition can be selected from cancer, rheumatoid arthritis,

AIDS, coronary heart disease, dehydration, malnutrition, lead exposure, malaria, respiratory disease, old age, 20 hypothyroidism, tuberculosis, hypopituitarism, neurasthenia, hypernatremia, hyponatremia, kidney disease, splenic, ankylosing spondylitis, decline (growth retardation) syndrome or any combination thereof.

In an embodiment of the invention, the method may include administration of a factor antagonist associated with

Dt LA Frtüru.OAO INDUSTRIAL cachexia, a factor associated with weakness, a factor associated with fatigue and / or a factor associated with fever. Factor associated with cachexia, factor associated with weakness, factor associated with fatigue and / or factor associated with fever can be selected from tumor necrosis factor alpha, gamma interferon, interleukin 1 alpha, interleukin 1 beta, interleukin 6 , factor that induces proteolysis, factor that inhibits leukemia or any combination thereof.

In one embodiment of the invention, the method may include the administration of an anti-cachexia agent selected from cannabis, dronabinol (Marinol ™), nabilone (Cesamet), cannabidiol, cannabicromeno, tetrahydrocannabinol, Sativex, megestrol acetate or any combination of the themselves.

In one embodiment of the invention, the method may include the administration of an antinausea or antiemetic agent selected from 5-HT3 receptor antagonists, ajwain, alizapride, anticholinergics, antihistamines, aprepitant, benzodiazepines, cannabicromeno, cannabidiol, cannabinoids, cannabis, casopitant, chlorpromazine, cyclizine, dexamethasone, dexamethasone, dimenhydrinate (Gravol ™), diphenhydramine, dolasetron, domperidone, dopamine antagonists, doxylamine, dronabinol (Marinol1”)1, rfroperidol, emetrol, ginger, granisetron, haloperidol, hydroxyzine, hyoscine, lorazepam, meclizine, metoclopramide, midazolam, muscimol, nabilone

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MEXICAN INDUSTRIAL PROPERTY INSTITUTE (Cesamet), nkl receptor antagonists, ondansetron, palonosetron, peppermint, Fenergam, prochlorperazine, Promacot, promethazine, Pentazine, propofol, sativex, tetrahydrocannabinol, trimethobenzamide, tropisetron, ghrelin, myostatin antagonists, anti-myostatin antibodies, selective androgen receptor modulators, selective estrogen receptor modulators, angiotensin AII antagonists, beta 10 adrenergic receptor agonists, beta three adrenergic receptor agonists, or any combination thereof.

In one embodiment of the invention, the patient's fever can be assessed by measuring the patient's body temperature.

In an embodiment of the invention, the method may include measuring the patient's body temperature prior to administration of the anti-IL-6 antibody and administering the anti-IL-6 antibody or antibody fragment if the patient's body temperature is greater than about

38 ° C.

In one embodiment of the invention, the method may include measuring the patient's body temperature within the range of anti-IL-6 antibody administration and administering the anti-IL-6 antibody or antibody fragment. 6 if the patient's body temperature indicates the

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presence of fever.

In one embodiment of the invention, the method may further include measuring the patient's body weight prior to administration of the anti-IL-6 antibody and administering the anti-IL6 antibody or antibody fragment if the patient's weight decreased by more than about 5% within about 30 days or if the patient's lean body mass index is less than about 17 kg / m2 (male patient) or less at about 14 kg / m2 (female patient).

In an embodiment of the invention, the method may include measuring the patient's muscle strength prior to administration of the anti-IL-6 antibody and administering the anti-IL-β antibody or antibody fragment if the patient's muscle strength decreased in a percentage greater than approximately 20% within approximately 30 days.

In one embodiment of the invention, the method can result in a prolonged improvement of the patient's cachexia, weakness, fatigue and / or fever.

In one embodiment of the invention, the patient's body mass can be increased by about 1 kilogram within about 4 weeks from administration of the anti-IL-6 antibody or antibody fragment.

In one embodiment of the invention, the patient's cachexia can be measurably improved within

approximately 4 weeks after administration of the anti-IL-6 antibody.

In one embodiment of the invention, the patient's cachexia can be evaluated by measuring the patient's total body mass, lean body mass, lean body mass index, and / or appendicular lean body mass.

In an embodiment of the invention, the measurement of the patient's body mass can discount (subtract) the estimated weight of the patient's tumor (s) and / or the extravascular fluid collections.

In one embodiment of the invention, the patient's cachexia can be measurably improved approximately 8 weeks after administration of the anti-IL-6 antibody.

In one embodiment of the invention, patient weakness can be measurably improved within approximately 4 weeks from administration of the anti-IL-6 antibody.

<td>In</td><td>a</td><td>modality of</td><td>the</td><td>invention, weakness</td><td>of the</td>

<td>patient</td><td>I know</td><td colspan="3">can measure by force test</td><td>of</td>

<td>Pressure.</td><td></td><td></td><td></td><td></td><td></td>

<td>In</td><td>a</td><td>modality of the</td><td colspan="2">invention the pressure force</td><td>of the</td>

<td>patient</td><td colspan="2">can be improved on</td><td>to the</td><td>minus about 15</td><td>% o</td>

<td>at least</td><td colspan="3">about 20%.</td><td></td><td></td>

<td>In</td><td>a</td><td>modality of</td><td>the</td><td>invention, weakness</td><td>of the</td>

Patient can be measurably improved approximately 8 weeks after administration of anti-IL-6 antibody.

In one embodiment of the invention, patient fatigue can be measurably improved within approximately 1 week from administration of the anti-IL-6 antibody.

In one embodiment of the invention, patient fatigue can be measured by the FACIT-F FS test.

In one embodiment of the invention, the patient's FACITF FS score can be improved by at least about points.

In one embodiment of the invention, patient fatigue can be measurably improved approximately 8 weeks after administration of the anti-IL-6 antibody.

In one embodiment of the invention, the patient's fever can be measurably improved within approximately 1 week from administration of the anti-IL-6 antibody.

In one embodiment of the invention, the patient's fever can be measurably improved approximately 8 weeks after administration of the anti-IL-6 antibody.

In one embodiment of the invention, the survivability of the patient can be improved.

In an embodiment of the invention, the quality of life of the patient can be improved.

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O £ LA INDUSTRIAL i'AOPIfDAO

BRIEF DESCRIPTION OF THE DIFFERENT VIEWS OF THE DRAWINGS

Fig. 1 shows that a variety of unique epitopes were recognized by harvesting anti-IL-6 antibodies prepared by the antibody selection protocol. The variability of epitopes was confirmed by means of anti-IL-6 antibody binding competition studies (ForteBio

Octet).

Fig. 2 shows the alignments of variable light and heavy chain sequences between rabbit antibody variable heavy and variable light chain sequences and homologous human sequences and humanized sequences. The flanking regions are identified as FR1-FR4. Complementarity determining regions are identified as CDR1CDR3. Amino acid residues are numbered as shown. The initial rabbit sequences are named RbtVL and RbtVH for the variable heavy and light chain sequences respectively. Three of the most similar human germ antibody sequences, spanning from Flanking Region 1 to the end of Flanking Region 3, are aligned below the rabbit sequences. The human sequence considered the most similar to the rabbit sequence is shown first. In the present example, those most similar sequences are L12A for the light chain and 3-6404 for the heavy chain. CDR3 sequences not shown

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INDUSTRIAL human. The closest human Flanking Region 4 sequence is aligned below the rabbit Flanking Region 4 sequence. The vertical dashes indicate a residue where the rabbit residue is identical to one or more of the human residues at the same position. Residues in bold indicate that the human residue at that position is identical to the rabbit residue at the same position. The humanized end sequences are named VLh and VHh for the variable light and heavy chain sequences respectively. Underlined residues indicate that the residue is equal to the rabbit residue at that position but different than the human residues at that position in the three aligned human sequences.

Fig. 3 shows the high correlation between produced IgG and antigen specificity for an IL-6 protocol example. 9 out of 11 wells showed specific IgG correlation with antigen recognition.

Fig. 4 provides the dose response curve of oi-2-macroglobulin (A2M) for Abl antibody administered intravenously at different doses one hour after a dose of human IL-6 of 100 pg / kg sc

Fig. 5 provides survival data for Abl antibody progression groups against control groups.

Fig. 6 provides additional survival data.

for the regression groups of the Abl antibody against the control groups.

Fig. 7 provides survival data for polyclonal human IgG at 10 mg / kg iv every three days (tumor size 270-320 mg) against Abl antibody at 10 mg / kg iv

every three days (tumor size 270-320 mg).

Fig. 8 provides survival data for polyclonal human IgG at 10 mg / kg iv every three days (tumor size 400-527 mg) against Abl antibody at 10 mg / kg iv every three days (tumor size 400 -527 mg).

Fig. 9 provides a pharmacokinetic profile of the Abl antibody in macaque monkeys. Plasma Abl antibody levels were quantified by antigen capture ELISA. This protein shows a half-life of 12-17 days that is consistent with other full-length humanized antibodies.

Fig. 10 (AD) provides binding data for the Ab4, Ab3, Ab8 and Ab2 antibodies, respectively. Fig. 10 E provides binding data for the Abl, Ab6 and Ab7 antibodies.

Fig. 11 summarizes the binding data of Fig. 10 (AE) in tabular form.

Fig. 12 presents the sequences of the 15 amino acid peptides used in the peptide mapping experiment of Example 14.

Fig. 13 presents the results of the bands

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prepared in Example 14.

Fig. 14 presents the results of the bands prepared in Example 14.

Fig. 15A shows the affinity and binding kinetics of

Abl to IL-6 of various species.

Fig. 15B shows the inhibition of IL-6 by Abl in the T1165 cell proliferation assay.

Fig. 16 shows the average plasma Abl concentration resulting from the single administration of Abl to healthy male subjects in various dose groups.

Fig. 17 shows the average area under the plasma Abl concentration time curve (AUC) for the dose groups in Fig. 16.

<td>Fig.</td><td> 18</td><td>shows</td><td>the</td><td colspan="2">average peak concentration</td><td>of</td>

<td>Abl in plasma</td><td colspan="2">(Cmax) for</td><td>the</td><td>dose groups of Fig.</td><td> 16.</td><td></td>

<td>Fig.</td><td> 19</td><td colspan="2">summarizes the</td><td>pharmacokinetic measurements</td><td>of</td><td>Abl</td>

<td>of the groups</td><td>of</td><td>dose of</td><td>the</td><td>Fig. 16.</td><td></td><td></td>

<td>Fig.</td><td> 20</td><td>shows</td><td>the</td><td>average concentration of</td><td>Abl</td><td>in</td>

<td>plasma that</td><td colspan="2">results from</td><td>the</td><td>single administration of</td><td>Abl</td><td>to</td>

patients with advanced cancer.

Fig. 21 illustrates the unprecedented elimination half-life of Abl compared to other anti-IL-6 antibodies.

Fig. 22 shows the increase in hemoglobin concentration after Abl administration to patients with

advanced cancer.

Fig. 23 shows the average plasma lipid concentrations after Abl administration to patients with advanced cancer.

Fig. 24 shows average neutrophil counts after Abl administration to patients with advanced cancer.

Fig. 25 shows the suppression of serum CRP levels in healthy individuals.

Fig. 26 (AB) shows the suppression of the levels of

Serum CRP in patients with advanced cancer.

Fig. 27 shows the prevention of weight loss by Abl in a mouse cancer cachexia model.

Fig. 28 shows the physical appearance of representative Abl-treated and control mice in the cancer cachexia model.

Fig. 29 shows that Abl promotes weight gain in patients with advanced cancer.

Fig. 30 shows that Abl decreases fatigue in patients with advanced cancer.

Fig. 31 shows that Abl promotes grip strength in patients with advanced cancer.

Fig. 32 shows that Abl suppresses an acute phase protein (serum amyloid A) in mice.

Fig. 33 shows that Abl increases the concentration of

plasma albumin in patients with advanced cancer.

Figs. 34 and 35 show alignments between variable heavy and light rabbit antibody sequences and homologous human sequences and the final humanized sequences. The flanking regions are identified as FR1-FR4.

Complementarity determining regions are identified as CDR1-CDR3.

Figs. 36 and 37 show alignments between the variable heavy and light sequences, of different forms of

Abl, respectively. The flanking regions are identified as FR1-FR4. Complementarity determining regions are identified as CDR1-CDR3. The differences in the sequences within the CDR regions are highlighted.

<td>Fig. 38</td><td>shows</td><td>than</td><td>the</td><td colspan="2">Abl increases hemoglobin</td>

<td>average at 80,</td><td>160 and</td><td> 320</td><td>mg</td><td>after 12 weeks</td><td>of</td>

<td>dosage.</td><td></td><td></td><td></td><td></td><td></td>

<td>The figure</td><td colspan="2">39 shows</td><td>the</td><td>average change from</td><td>the</td>

Reference hemoglobin for the data presented in Figure 38.

Figure 40 shows that Abl increases the average hemoglobin to 160 and 320 mg after 12 weeks of dosing in patients who have the reference hemoglobin below 11 g / 1.

Fig. 41 shows that Abl increases the average hemoglobin to 80, 160 and 320 mg after 16 weeks of

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dosage.

Figure 42 shows the data of change in the averaged weight of each dose concentration group (placebo, 80 mg, 160 mg, and 320 mg) of the monoclonal antibody Abl over 12 weeks.

Fig. 43 shows the averaged percentage change in body weight of each dose concentration group corresponding to Fig. 42.

Figure 44 shows the change in the average lean body mass data for the dose concentration groups corresponding to Figure 42.

Figure 45 shows increases in the average Facit-F FS subscale score for some of the dose concentration groups in the patient population after dosing of 80, 160, and 320 mg after 8 weeks.

Fig. 46 shows the change in the Facit-F FS subscale score from the reference point corresponding to Fig. 45.

DETAILED DESCRIPTION OF THE PREFERRED MODALITIES

I defined gifts

It should be understood that the present invention is not limited to the particular methodology, protocols, cell lines, species or animal genera and reagents described, since the

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY may vary. Terminology used herein is also to be understood for the purpose of describing only particular embodiments and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

As used herein the singular forms un / a, and and the include plural referents unless the context clearly states otherwise. Accordingly, for example, reference to a cell includes a plurality of such cells, and reference to protein includes reference to one or more proteins and equivalents thereof known to those skilled in the art, and so on. All technical and scientific terms used herein have the same meaning as understood by the person skilled in the art to which the present invention belongs unless otherwise indicated.

Interleukin-6 (IL-6): As used herein, Interleukin-6 (IL-6) comprises not only the following 212 amino acid sequence available as access to GenBank protein No. NP_000591:

MNSFSTSAFGPVAFSLGLLLVLPAAFPAPVPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDG

ISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYL

EYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDM

TTHLILRSFKEFLQSSLRALRQM (SEQ ID NO: 1), but also any pre-pro, pro-, and mature forms of this amino acid sequence

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of IL-6, as well as mutants and variants that include allelic variants of this sequence.

Disease or condition: As used herein, disease or condition refers to a disease or condition that a patient has been diagnosed with or suspected of having, particularly a disease or condition associated with

IL-6 elevated. A disease or condition comprises, without limitation, the side effects of medications or treatments (such as radiation therapy), as well as idiopathic conditions characterized by symptoms including elevated IL-6.

Cachexia: As used herein, cachexia, also known as wasting disease, refers to any disease marked especially by progressive emaciation, weakness, general unhealthiness, malnutrition, loss of body mass, loss of muscle mass, or an accelerated loss of skeletal muscle in the context of a chronic inflammatory response (discussed in Kotler, Ann Intern

Med. 2000 Oct 17; 133 (8): 622-34). The diseases and conditions in which cachexia is frequently observed include cancer, rheumatoid arthritis, AIDS, coronary heart disease, dehydration, malnutrition, lead exposure, malaria, respiratory disease, old age, hypothyroidism, tuberculosis, hypopituitarism, neurasthenia, hypernatremia, hyponatremia, kidney disease, splenic, ankylosing spondylitis,

decline syndrome (growth retardation) and other diseases, particularly chronic diseases. Cachexia can also be idiopathic (arising from an uncertain cause). Assessment of weight in a patient is understood to exclude tumors or fluid accumulation, eg, tumor weight, accumulation of extravascular fluid, etc. Cachexia can be assessed by measuring the patient's total body mass (except for tumors or fluid accumulation), total lean mass (without fat), body mass, lean mass of arms and legs (lean appendicular mass, eg, measured using dual energy x-ray absorptiometry or bioelectrical impedance spectroscopy), and / or lean body mass index (lean body mass divided by the height of the patient squared). See Kotler, Ann Intern Med. 2000 Oct 17; 133 (8): 622-34; Marcora et al., Rheumatology (Oxford).

2006 Nov; 45 (ll): 1385-8.

Weakness: As used herein, weakness refers to physical fatigue, which typically manifests as a loss of strength and / or muscular endurance. Weakness can be central (affecting most or all of the body's muscles) or peripheral (affecting a subset of muscles). Weakness includes actual weakness in which the patient's muscles decrease to some extent the production of peak and / or sustained force, and perceived weakness, in which the patient perceives that more effort is required to

perform a task even though the force measured objectively remains almost the same, and the patient can measure it objectively or report it himself. For example, weakness can be measured objectively using the grip strength test (a medically recognized test to assess muscle strength), typically using a grip dynamometer.

Fatigue·. As used herein, fatigue refers to mental fatigue (for physical fatigue see weakness). Fatigue includes drowsiness (drowsiness) and / or decreased attention. Fatigue can be measured using a variety of tests known in the art, such as the FACIT-F test (Functional Assessment of Chronic Disease Treatment - Fatigue). See, for example, Celia, D., Lai, JS, Chang, CH, Peterman, A., & Slavin, M. (2002). Fatigue in cancer patients compared with fatigue in the general population. Cancer, 94 (2),

528-538; Celia, D., Eton, DT, Lai, F JS., Peterman, AH &

Merkel, DE (2002). Combining anchor and distribution based methods to derive minimal clinically important differences on the Functional Assessment of Cancer Therapy anemia and fatigue scales. Journal of Pain & Symptom Management, 24 (6) 547-561.).

Fever·. As used herein, fever refers to the reference point of body temperature that increases by at least 1 to 2 degrees Celsius. Fever is commonly associated with a subjective feeling of hypothermia that manifests

as a cold sensation, tremors, increased heart rate and respiratory rate so that the individual's body reaches the increased benchmark. As understood in medicine, normal body temperature typically varies with activity level and time of day, with maximum temperatures observed in the afternoon and before night, and minimum temperatures observed during the second half of the sleep, and temperature measurements may be influenced by external factors such as mouth breathing, consumption of food or drink, smoking, or room temperature (depending on the type of measurement). Also, the normal temperature reference point for individuals can vary up to around 0.5 degrees Celsius, therefore an individual's temperature can be interpreted by a medical professional based on these factors to diagnose if they have fever. Fever is typically diagnosed by an internal body temperature above 38.0 degrees Celsius, an oral temperature above 37.5 degrees Celsius, or an axillary temperature above 37.2 degrees Celsius.

Improved'. As used herein, improved, improved, and other grammatical variants include any beneficial changes resulting from treatment. A beneficial change is any way in which the patient's condition is better than it would be in the absence of treatment.

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Enhanced includes preventing an unwanted condition, slowing the rate of worsening of the condition, delaying the development of an unwanted condition, and restoring an essentially normal state. For example, cachexia improvement includes any increase in patient mass, such as total body mass (excluding weight that is normally excluded during cachexia evaluation, eg, tumor weight, accumulation of extravascular fluid, etc.), lean body mass and / or lean appendicular mass, as well as any delay or slowdown in the rate of mass loss, or prevention or slowdown of loss of mass associated with a disease or condition that the patient was diagnosed with. For another example, improvement in weakness includes any increase in the patient's strength, as well as any delay or slowdown in the rate of loss of strength or prevention or slowdown of loss of strength associated with a disease or condition the patient was diagnosed with. . As a further example, improvement in fatigue includes any decrease in patient fatigue, as well as any delay or slowdown in the rate of increased fatigue or prevention or slowdown of increased fatigue associated with a diagnosed disease or condition to the patient. As a further example, improvement in fever includes any decrease in the patient's fever, as well as any delay or slowdown

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of the increased fever associated with a disease or condition the patient was diagnosed with.

C-reactive protein (CRP): As used herein, C-reactive protein (CRP) not only comprises the following 224 amino acid sequence available as GenBank Access No. to protein NP_000558:

MEKLLCFLVLTSLSHAFGQTDMSRKAFVFPKESDTSYVSLKAPLTKPLKAFTVCLHFYTELSS

TRGYSIFSYATKRQDNEILIFWSKDIGYSFTVGGSEILFEVPEVTVAPVHICTSWESASGIVE

FWVDGKPRVRKSLKKGYTVGAEASIILGQEQDSFGGNFEGSQSLVGDIGNVNMWDFVLSPDEI

NTIYLGGPFSPNVLNWRALKYEVQGEVFTKPQLWP (SEQ ID NO: 726), but also any pre-pro, pro-, and mature forms of this amino acid sequence of CRP, as well as mutants and variants including allelic variants of this sequence. CRP levels, eg, in serum, liver, tumor, or elsewhere in the body, can be easily measured using routine methods and commercially available reagents, eg. ELISA, antibody test strip, immunoturbidimetry, rapid immunodiffusion, visual agglutination, Western blot,

Northern blot etc.

Interleukin-6 receptor (IL-6R), also called IL-6 alpha receptor (IL-6RA): As used herein, interleukin-6 receptor (IL-6R, also IL-6 alpha receptor or IL-6RA) not only comprises the following 468 amino acid sequence available as Accession No.

Swiss-Prot la

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P08887:

MLAVGCALLAALLAAPGAALAPRRCPAQEVARGVLTSLPGDSVTLTCPGVEPEDNATVHWVLR

KPAAGSHPSRWAGMGRRLLLRSVQLHDSGNYSCYRAGRPAGTVHLLVDVPPEEPQLSCFRKSP

LSNVVCEWGPRSTPSLTTKAVLLVRKFQNSPAEDFQEPCQYSQESQKFSCQLAVPEGDSSFYI

VSMCVASSVGSKFSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTWQDPHSWNSSFYRLR

FELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHWQLRAQEEFGQGEWSEWSPEAMGTPW

TESRSPPAENEVSTPMQALTTNKDDDNILFRDSANATSLPVQDSSSVPLPTFLVAGGSLAFGT

LLCIAIVLRFKKTWKLRALKEGKTSMHPPYSLGQLVPERPRPTPVLVPLISPPVSPSSLGSDN

TSSHNRPDARDPRSPYDISNTDYFFPR (SEQ ID NO: 727), but also any pre-pro, pro-, and mature forms of this amino acid sequence, as well as mutants and variants including allelic variants of this sequence.

gpl30: As used herein, gpl30 (also called interleukin-6 beta subunit receptor) not only comprises the following 918 amino acid precursor sequence available as Swiss-Prot Access No. to Protein:

P40189:

MLTLQTWVVQALFIFLTTESTGELLDPCGYISPESPVVQLHSNFTAVCVLKEKCMDYFHVNAN

YIVWKTNHFTIPKEQYTIINRTASSVTFTDIASLNIQLTCNILTFGQLEQNVYGITIISGLPP

EKPKNLSCIVNEGKKMRCEWDGGRETHLETNFTLKSEWATHKFADCKAKRDTPTSCTVDYSTV

YFVNIEVWVEAENALGKVTSDHINFDPVYKVKPNPPHNLSVINSEELSSILKLTWTNPSIKSV

IILKYNIQYRTKDASTWSQIPPEDTASTRSSFTVQDLKPFTEYVFRIRCMKEDGKGYWSDWSE

EASGITYEDRPSKAPSFWYKIDPSHTQGYRTVQLVWKTLPPFEANGKILDYEVTLTRWKSHLQ

NYTVNATKLTVNLTNDRYLATLTVRNLVGKSDAAVLTIPACDFQATHPVMDLKAFPKDNMLWV

EWTTPRESVKKYILEWCVLSDKAPCITDWQQEDGTVHRTYLRGNLAESKCYLITVTPVYADGP

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GSPESIKAYLKQAPPSKGPTVRTKKVGKNEAVLEWDQLPVDVQNGFIRNYTIFYRTIIGNETA

VNVDSSHTEYTLSSLTSDTLYMVRMAAYTDEGGKDGPEFTFTTPKFAQGEIEAIVVPVCLAFL

LTTLLGVLFCFNKRDLIKKHIWPNVPDPSKSHIAQWSPHTPPRHNFNSKDQMYSDGNFTDVSV

VEIEANDKKPFPEDLKSLDLFKKEKINTEGHSSGIGGSSCMSSSRPSISSSDENESSQNTSST

VQYSTVVHSGYRHQVPSVQVFSRSESTQPLLDSEERPEDLQLVDHVDGGDGILPRQQYFKQNC

SQHESSPDISHFERSKQVSSVNEEDFVRLKQQISDHISQSCGSGQMKMFQEVSAADAFGPGTE

GQVERFETVGMEAATDEGMPKSYLPQTVRQGGYMPQ (SEQ ID NO: 728), but also any pre-pro, pro-, and mature forms of this amino acid sequence, such as the mature form encoded by amino acids 23-918 of the sequence shown, as well as mutants and variants that they include allelic variants of this sequence.

Glasgow Prognosis Scale {GPS): As used herein, the Glasgow Prognosis Scale (GPS) refers to an inflammation-based value that gives a prognostic point at a serum albumin level of less than <35 mg / L and one point at a CRP level above 10 mg / L. Therefore, a GPS of 0 indicates normal albumin and CRP, a GPS of 1 indicates less albumin or more CRP than normal, and a GPS of 2 indicates less albumin and more CRP than normal.

Effective amount: As used herein, effective amount, effective amount for, effective amount of X for and the like, refers to an amount of an active ingredient effective to alleviate or reduce to some extent one or more of the symptoms of the disease that needs treatment or

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to delay the onset of symptoms or clinical markers of a disease that needs prevention, when the compound is administered. Accordingly, an effective amount refers to an amount of the active ingredient that has effects such as (i) reversing the rate of progression of a disease, (ii) to some extent inhibiting further progression of the disease, and / or (iii) alleviate to some extent (or preferably eliminate) one or more symptoms associated with the disease. The effective amount can be empirically determined by experimenting with the compounds involved in known in vivo and in vitro model systems for a disease in need of treatment. The context in which the phrase effective amount is used may indicate a particular desired effect. For example, an amount of an anti-IL-6 antibody effective in reducing weakness and similar phrases refer to an amount of anti-IL-6 antibody that, when administered to a subject, causes a measurable decrease in weakness based on determined by the grip strength test. Similarly, an amount of an anti-IL-6 antibody effective in increasing weight and similar phrases refer to an amount of anti-IL-6 antibody that, when administered to a subject, causes a measurable increase in weight. of the patient. An effective amount will vary based on the individual's weight, sex, age, and medical history, as well as the severity of the patient's condition (s), type of disease, or

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diseases, mode of administration and the like. An effective amount can be easily determined using routine experiments, for example, by titration (administration of increasing doses until an effective dose is found) and / or reference to amounts that were effective in previous patients. In general, the anti-IL-6 antibodies of the present invention will be administered in doses ranging from about 0.1 mg / kg to about 20 mg / kg of the patient's body weight.

Prolonged improvement in cachexia: As used herein, protracted improvement in cachexia refers to a measurable improvement in the patient's body mass, lean body mass, appendicular lean body mass and / or lean body mass index. , relative to baseline (i.e., level prior to treatment initiation) that can be detected within approximately 4 weeks and remains improved for an extended period, for example at least around

<td>of</td><td> 35</td><td>days,</td><td>at least</td><td>around</td><td>of</td><td>40 days, at least</td><td>around</td>

<td>of</td><td> 50</td><td>days,</td><td>at least</td><td>around</td><td>of</td><td>60 days at least</td><td>around</td>

<td>of</td><td> 70</td><td>days,</td><td colspan="3">at least around</td><td>11 weeks or</td><td>at least</td>

about 12 weeks from the start of treatment

Long-term improvement in weakness: As used herein, long-term improvement in weakness refers to a measurable improvement in muscle strength, relative to baseline (i.e., the level prior to treatment initiation)

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which can be detected within about 2 weeks and remains improved for a long period, for example

<td>to the</td><td>less</td><td>around</td><td>of</td><td> 21</td><td>days,</td><td>to the</td><td>less</td><td>around</td><td>of</td><td> 28</td><td>days,</td>

<td>to the</td><td>less</td><td>around</td><td>of</td><td> 35</td><td>days,</td><td>to the</td><td>less</td><td>around</td><td>of</td><td> 40</td><td>days,</td>

<td>to the</td><td>less</td><td>around</td><td>of</td><td> 50</td><td>days,</td><td>to the</td><td>less</td><td>around</td><td>of</td><td> 60</td><td>days,</td>

at least around 70 days, at least around 11 weeks or at least around 12 weeks from the start of treatment.

Prolonged improvement in fatigue: As used herein, Prolonged improvement in fatigue refers to a measurable improvement in fatigue, relative to the baseline level (i.e. the level before the start of treatment) that can be detected during about 1 week and remains improved for an extended period, for example at least about 14 days, at least about 21 days, at

<td>less</td><td>around</td><td>of</td><td> 28</td><td>days,</td><td>to the</td><td>less</td><td>around</td><td>of</td><td> 35</td><td>days,</td><td>to the</td>

<td>less</td><td>around</td><td>of</td><td> 40</td><td>days,</td><td>to the</td><td>less</td><td>around</td><td>of</td><td> 50</td><td>days,</td><td>to the</td>

<td>less</td><td>around</td><td>of</td><td> 60</td><td>days,</td><td>to the</td><td>less</td><td>around</td><td>of</td><td> 70</td><td>days,</td><td>to the</td>

less around 11 weeks or at least around 12 weeks from the start of treatment.

Prolonged improvement in fever: As used herein, prolonged improvement in fever refers to a measurable decrease in fever (for example, the peak temperature or the amount of time the temperature remains high), relative to the starting level (i.e. the level before

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INSTITUTO MEXICANO D £ LA PROPERTY INDUSTRIAL from the start of treatment) that can be detected within approximately 1 week and remains improved for a

<td>prolonged period,</td><td>for example,</td><td>at least</td><td>around</td><td>of</td><td> 14</td><td>days,</td>

<td>at least around</td><td>21 days,</td><td>at least</td><td>around</td><td>of</td><td> 28</td><td>days,</td>

<td>at least around</td><td>35 days,</td><td>at least</td><td>around</td><td>of</td><td> 40</td><td>days,</td>

<td>at least around</td><td>50 days,</td><td>at least</td><td>around</td><td>of</td><td> 60</td><td>days,</td>

at least around 70 days, at least around 11 weeks, or at least around 12 weeks from the start of treatment.

Yeast Species That Can Pair: The present invention aims to broadly understand any diploid or tetraploid yeast that can grow in culture. Such yeast species can exist in haploid, diploid, or tetraploid form. The cells of a given ploidy can, under appropriate conditions, proliferate in this way for an undetermined number of generations. Diploid cells can also sporulate to form haploid cells. Sequential pairing can result in tetraploid strains by additional pairing or fusion of diploid strains. In the present invention, diploid or polyploid yeast cells are preferably produced by mating or fusion of spheroplasts. In one embodiment of the invention, the mating yeast is a member of the Saccharcmycetaceae family, which includes the genera Arxiozyma, Ascobotryozyma, Citeromyces, Debaryomyces, Dekkera, .a. Hee r

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Eremothecium, Issatchenkia, Kazachstania, Kluyveromyces,

Kodamaea, Lodderomyces, Pachysolen, Pichia, Saccharomyces, Saturnispora, Tetrapisispora, Torulaspora, Willíopsis and Zygosaccharomyces. Other types of yeast potentially useful in the invention include Yarrowia, Rhodosporidium, Candida, Hansenula, Filobasium, Filobasidellla, Sporidiobolus, Bullera, Leucosporidium, and Filobasidella.

In a preferred embodiment of the invention, the matable yeast is a member of the genus Pichia. In a further preferred embodiment of the invention, the mating yeast of the genus Pichia is one of the following species: Pichia pastoris, Pichia methanolica and Hansenula polymorpha (Pichia angusta). In a particularly preferred embodiment of the invention, the mating yeast of the genus Pichia is the species Pichia pastoris.

Haploid yeast cell: A cell that has a unique copy of each gene for its normal genomic (chromosomal) complement.

Polyploid yeast cell: A cell that has more than one copy of its normal genomic (chromosomal) complement.

Diploid Yeast Cell: A cell that has two copies (alleles) of essentially every gene in its normal genomic complement, typically formed by the fusion (mating) process of two haploid cells.

Tetraploid yeast cell: Cell that has four

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Copies (alleles) of essentially each gene from its normal genomic complement, typically formed by the fusion (pairing) process of two haploid cells. Tetraploids can have two, three, four, or more different expression cassettes. Such tetraploids could be obtained from S.

cerevisiae by selective pairing of heterozygous a / a and alpha / alpha homozygous diploids and in Pichia by sequential haploid pairing to obtain auxotrophic diploids. For example, a haploid [met his] can be paired with a haploid [ade his] to obtain a diploid [his] and a haploid [met arg] can be paired with a haploid [ade arg] to obtain a diploid [arg] and then diploid [his] x diploid [arg] to obtain a tetraploid prototroph. Those of skill in the art will understand that the reference to the benefits and uses of diploid cells may also apply to tetraploid cells.

Yeast pairing: The process by which two haploid yeast cells naturally fuse to form a diploid yeast cell.

Meiosis: The process by which a diploid yeast cell undergoes reductive division to form four haploid spore products. Each spore can then germinate and form a | vegetatively growing haploid cell line.

Select marker: A selectable marker is a

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gene or gene fragment that confers a growth phenotype (physical growth characteristic) to a cell that receives that gene, for example, through a transformation event. The selectable marker allows the cell to survive and grow in a selective growth medium under conditions in which cells that do not receive said selectable marker gene cannot grow. Selectable marker genes are generally comprised of several types, including positive selectable marker genes, such as a gene that confers resistance to an antibiotic or other drug on a cell, temperature when two ts mutants are crossed, or when a ts mutant is transformed; negative selectable marker genes, such as a biosynthetic gene that confers on a cell the ability to grow in a medium without a specific nutrient needed by all cells that do not have such a biosynthetic gene or a mutagenized biosynthetic gene that confers inability on a cell of growing by cells that do not have the wild type gene and the like. Suitable markers include but are not limited to: ZEO, G418, LYS3, METI, MET3a, ADE1, ADE3,

URA3 and the like.

Expression vector: These DNA vectors contain elements that facilitate manipulation for the expression of a foreign protein within the target host cell. Conveniently, sequence manipulation and production | Ρ ί κ j *. to

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INDUSTRIAL DNA for transformation is first carried out in a bacterial host, for example, E. coli and, in general, vectors will include sequences to facilitate such manipulations, including a bacterial origin of replication and a selection marker appropriate bacterial.

Selection markers encode proteins necessary for the survival or growth of transformed host cells grown in a selective culture medium. Host cells not transformed with the vector containing the selection gene will not survive in the culture medium. Typical selection genes encode proteins that (a) provide resistance to antibiotics or other toxins, (b) supplement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media. Examples of yeast transformation vectors and methods are described, for example, in Burke,

D., Dawson, D., & Stearns, T. (2000). Methods in yeast genetics: a Coid Spring Harbor Laboratory course manual.

Plainview, NY: Coid Spring Harbor Laboratory Press.

Expression vectors for use in methods of the invention will additionally include yeast-specific sequences including an auxotrophic or selectable drug marker to identify the transformed yeast strains. A drug marker can additionally be used to extend the copy number of the vector by

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY a yeast host cell.

The sequence of interest encoding the polypeptide is operably linked to transcriptional and translational regulatory sequences that provide for expression of the polypeptide in yeast cells. These vector components may include, but are not limited to, one or more of the following: an enhancer element, a promoter, and a transcription termination sequence. Sequences may also be included for secretion of the polypeptide, eg, a signal sequence and the like.

A yeast origin of replication is optional, since expression vectors are commonly integrated into the yeast genome.

In one embodiment of the invention, the polypeptide of interest is operably linked, or fused, to sequences that provide optimized secretion of the polypeptide from diploid yeast cells.

Nucleic acids are operably linked when they are placed into a functional relationship with another nucleic acid sequence. For example, DNA from a signal sequence is operably linked to DNA from a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence. Generally, operationally. UT 'I

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<td>bound</td><td colspan="2">means that</td><td>the sequences</td><td>of</td><td>DNA that</td><td>is it so</td>

<td>linked</td><td>are</td><td>adjoining,</td><td>and, in the case of</td><td>a</td><td colspan="2">secretarial leader,</td>

<td>adjoining</td><td>and</td><td>in frame</td><td>Reading.</td><td>Without</td><td>embargo,</td><td>the</td>

<td colspan="2">enhancers</td><td colspan="2">they do not have to be contiguous.</td><td>The</td><td>link it</td><td>achieves</td>

by ligation at convenient restriction sites or, alternatively, by a PCR / recombination method known to those of skill in the art (Gateway * Technology; Invitrogen, Carlsbad California). If such sites do not exist, synthetic oligonucleotide linkers or adapters are used in accordance with conventional practice.

Promoters are untranslated sequences located upstream (5 ') of the initial codon of a structural gene (generally between about 100 and 1000 bp) that control the transcription and translation of particular nucleic acid sequences to which they are operably linked. Such promoters fall into several classes: inducible, constitutive, and repressible promoters (which increase transcription levels in response to the absence of a repressor). Inducible promoters can initiate increased levels of DNA transcription under their control in response to some change in culture conditions, for example, the presence or absence of a nutrient or a change in temperature.

The yeast promoter fragment may also serve as a site for homologous recombination and integration of the

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY expression vector at the same site in the yeast genome;

alternatively, a selectable marker is used as the site for homologous recombination. The Pichia transformation is described in Cregg et al. (1985) Mol. Cell. Biol.

5:3376-3385.

Examples of suitable Pichia promoters include the AOX1 promoter (Cregg et al. (1989) Mol. Cell. Biol. 9: 13161323), the ICL1 promoter (Menendez et al. (2003) Yeast

20 (13): 1097-108), glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter (Waterham et al. (1997) Gene 186 (1): 37-44) and FLD1 promoter (Shen et al. (1998) Gene 216 (1): 93-102). The GAP promoter is a strong constitutive promoter and the AOX and FLD1 promoters are inducible.

Other yeast promoters include ADH1, alcohol dehydrogenase II, GAL4, PHO3, PHO5, Pyk, and chimeric promoters derived therefrom. Additionally, non-yeast promoters such as mammalian, insect, plant, reptile, amphibian, viral, and bird promoters can be used in the invention. More typically, the promoter will comprise a mammalian promoter (potentially endogenous to the expressed genes) or comprise a yeast or viral promoter that provides effective transcription to yeast systems.

Polypeptides of interest can be produced recombinantly not only directly, but also as a

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fusion polypeptide with a heterologous polypeptide, eg, a signal sequence or other polypeptide that has a specific cleavage site at the N-terminus of the mature polypeptide or protein. In general, the signal sequence may be a component of the vector or it may be a part of the polypeptide coding sequence introduced into the vector. The selected heterologous signal sequence is preferably one that is recognized and processed through one of the available standard pathways within the host cell. The S. cerevisiae alpha factor pre-pro signal has proven efficacy in secreting a variety of recombinant P. pastoris proteins. Other yeast signal sequences include the alpha mating factor signal sequence, the invertase signal sequence, and signal sequences that come from other secreted yeast polypeptides. Additionally, these signal peptide sequences can be designed to provide enhanced secretion in diploid yeast expression systems. Other secretion signals of interest also include mammalian signal sequences, which may be heterologous to the protein being secreted or may be a native sequence for the protein being secreted. The signal sequences include prepeptide sequences and, in some cases, can include propeptide sequences. Many of such signal sequences are known in the art,

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including signal sequences found in immunoglobulin chains, eg, preprotoxin sequence K28, PHA-E, FACE, human MCP-1, albumin signal sequences in human serum, human Ig heavy chain, human Ig light chain and the like . For example, see Hashimoto et. to the. Protein Eng

11 (2) 75 (1998) and Kobayashi et. to the. Therapeutic Apheresis 2 (4)

257 (1998).

Transcription can be increased by inserting a transcription activating sequence into the vector. These activators are cis-acting elements of DNA, generally around 10 to 300 bp, that act on a promoter to increase its transcription. Transcription enhancers are relatively independent of orientation and position, having been found 5 'and 3' in relation to the transcription unit, within an intron, as well as within the coding sequence itself. The enhancer may be spliced into the expression vector at the 5 'or 3' position with respect to the coding sequence, but is preferably located at the 5 'site of the promoter.

Expression vectors used in eukaryotic host cells may also contain sequences necessary for termination of transcription and to stabilize mRNA. Such sequences are commonly available from 3 'to the translation stop codon, in untranslated regions of eukaryotic or viral DNA or cDNA. These regions contain

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Constructing suitable vectors containing one or more of the components listed above employs standard ligation techniques or PCR / recombination methods. Isolated plasmids or DNA fragments are excised, custom made, and religated as desired to generate the necessary plasmids or by recombination methods. For analysis to confirm correct sequences in constructed plasmids, ligation mixtures are used to transform host cells and successful transformers selected for resistance to an antibiotic (eg, ampicillin or Zeocin ™ (phleomycin)) where appropriate. Transformer plasmids are prepared, analyzed by restriction endonuclease digestion, and / or sequenced.

As an alternative to fragment restriction and ligation, recombination methods based on att sites and recombination enzymes can be used to insert DNA sequences into a vector. Such methods are described, for example, by Landy (1989) Ann.Rev.Biochem. 58: 913-949 and are known to those of skill in the art. These methods use intermolecular DNA recombination mediated by a mixture of recombination proteins encoded by lambda and E.coli. Recombination occurs between specific binding sites (att) on the interacting DNA molecules. For a description of

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY att sites see Weisberg and Landy (1983) Site-Specific

Recombination in Phage Lambda, in Lambda II, Weisberg, ed. (Coid Spring Harbor, NY: Cold Spring Harbor Press), pp. 211-250. The DNA segments flanking the recombination sites are changed so that after recombination, the att sites are hybrid sequences that comprise sequences donated by each parent vector. Recombination can occur between DNAs of any topology.

The att sites can be introduced into a sequence of interest by ligation of the sequence of interest into an appropriate vector, the generation of a PCR product containing att B sites using specific primers, the generation of a cloned cDNA library in an appropriate vector containing att sites and the like.

Folding, as used herein, refers to the three-dimensional structure of polypeptides and proteins, where interactions between amino acid residues act to stabilize the structure. While non-covalent interactions are important in determining structure, in general the proteins of interest will have intra- and / or intermolecular covalent disulfide bonds formed by two cysteine residues. For naturally-occurring proteins and polypeptides or derivatives and variants thereof, appropriate folding is typically the array that results in activity

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optimal biological and can be controlled1NUConvenience through activity assays, for example, binding of TigandoT enzyme activity, etc.

In some cases, for example when the desired product is of synthetic origin, tests based on biological activity will be less significant. The proper folding of such molecules can be determined based on physical properties, energy considerations, modeling studies, and the like.

The expression host can be further modified by introducing sequences that encode one or more enzymes that enhance folding and formation of disulfide bonds, i.e. foldases, chaperonins, etc. Such sequences can be expressed constitutively or inducible in the yeast host cell using vectors, markers, etc., as known in the art. Preferably, sequences, including transcriptional regulatory elements sufficient for the desired expression pattern, are stably integrated into the yeast genome through targeted methodology.

For example, eukaryotic PDI is not only an effective catalyst for cistern protein oxidation and disulfide binding isomerization, but it also exhibits chaperone activity. Co-expression of PDI can facilitate the production of active proteins that possess multiple binding of

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disulfide. Also of interest is the expression of BIP (immunoglobulin heavy chain binding protein), cyclophilin, and the like. In one embodiment of the invention, each of the haploid parent strains expresses a distinctive folding enzyme, for example, one strain can express BIP and the other strain can express PDI or combinations thereof.

The terms "desired protein" or target protein are used interchangeably and generally refer to a humanized antibody or a binding portion thereof described herein. The term antibody is intended to include any molecular structure that contains a polypeptide chain with a specific shape that is suitable for and recognizes an epitope, where one or more non-covalent binding interactions stabilize the complex between the molecular structure and the epitope. The archetypal antibody molecule is immunoglobulin and all types of immunoglobulins, IgG, IgM, IgA, IgE, IgD, etc., from all sources, eg, human, rodent, rabbit, cow, sheep, pig, dog , other mammals, chickens, other birds, etc., are considered antibodies. A preferred source for producing antibodies useful as a starting material according to the invention are rabbits. Numerous antibody coding sequences have been described, and others may arise by methods known in the art. Examples thereof include chimeric antibodies, human antibodies, and others.

non-human mammalian antibodies, humanized antibodies, single-chain antibodies such as scFvs, camelid antibodies, nano-antibodies, IgNAR (shark-derived single-chain antibodies), small molecule immunological pharmaceuticals (SMIP), and antibody fragments such as Fab , Fab ', F (ab') 2 and the like. See Streltsov VA, et al., Structure of a shark IgNAR antibody variable domain and modeling of an early-developmental isotype, Protein Sci.

November 2005; 14 (11): 2901-9. Epub 2005 Sep 30; Greenberg AS, et al., Ά new antigen receptor gene family that undergoes rearrangement and extensive somatic diversification in sharks,

Nature. March 9, 1995; 374 (6518): 168-73; Nuttall SD, et al., Isolation of the new antigen receptor from wobbegong sharks, and use as a scaffold for the display of protein loop librarles, Mol Immunol August 2001; 38 (4): 313-26; HamersCasterman C, et al., Naturally occurring antibodies devoid of light chains, Nature June 3, 1993; 363 (6428): 446-8; Gilí DS, et al., Biopharmaceutical drug discovery using novel protein scaffolds, Curr Opin Biotechnol. December 2006;

17 (6): 653-8. Electronic publication of October 19, 2006.

For example, antibodies or antigen-binding fragments can be produced by genetic engineering. In this technique, as with other methods, cells that produce antibodies are sensitized to the desired antigen or immunogen. Messenger RNA isolated from cells

Antibody-producing is used as a template to produce cDNA using PCR amplification. A vector library, each containing a heavy chain gene and a light chain gene that retain the initial antigen specificity, is produced by inserting appropriate sections of the amplified immunoglobulin cDNA into the expression vectors. A combinatorial library is constructed by combining the heavy chain gene library and the light chain gene library.

This results in a library of clones co-expressing a light and heavy chain (resembling either the Fab fragment or the antigen-binding fragment of an antibody molecule). Vectors carrying these genes are cotransfected in a host cell. When antibody gene synthesis is induced in the transfected host, the heavy and light chain proteins assemble themselves to produce active antibodies that can be detected by analysis with the antigen or immunogen.

Sequences of interest encoding antibodies include those encoded by native sequences, as well as nucleic acids which, due to the degeneracy of the genetic code, are not sequentially identical to the described nucleic acids and variants thereof. Variant polypeptides can include amino acid (aa) substitutions, additions, or deletions. Amino acid substitutions can be

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INDUSTRIAL conservative amino acid substitutions or substitutions to remove non-essential amino acids, such as to alter a glycosylation site or to minimize misfolding by substituting or removing one or more cysteine residues that are not necessary to function. Variants can be designed to retain or have enhanced biological activity of a particular region of the protein (eg, a functional domain, catalytic amino acid residues, etc.). Variants also include fragments of the polypeptides described herein, in particular biologically active fragments and / or fragments corresponding to functional domains. Techniques for in vitro mutagenesis of cloned genes are known. Also included in the subject invention are polypeptides that have been modified using common molecular biological techniques to improve their resistance to proteolytic degradation, to optimize solubility properties, or to make them more suitable as therapeutic agents.

Chimeric antibodies can be made through recombinant means by combining the variable light and heavy chain regions (VL and VH) obtained from cells that produce antibodies of one species with the light and heavy chain constant regions of others. Typically, chimeric antibodies use variable regions of rodents or rabbits and human constant regions to produce an antibody.

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY predominantly with human domains. The production of such chimeric antibodies is known in the art and can be accomplished by standard means (as described, for example, in US Patent No. 5,624,659, incorporated herein by reference in its entirety). Additionally, it is contemplated that the human chimeric antibody constant regions of the invention can be selected from the IgGl, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, constant regions.

IgG9, IgGlO, IgGll, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17,

IgG18 or IgG19.

Humanized antibodies are designed to contain even more human-type immunoglobulin domains and incorporate only the complementarity determining regions of the animal derived antibody. This is accomplished by carefully examining the sequence of the hypervariable loops of the variable regions of the monoclonal antibody and matching them to the structure of the human antibody chains. Although the process seems complex, it is simple in practice. See, for example, US Patent No. 6,187,287, incorporated herein by reference in its entirety.

In addition to complete immunoglobulins (or their recombinant equivalents), immunoglobulin fragments comprising the epitope binding site (eg, Fab ', F (ab') 2 or other fragments) can be synthesized. Fragments or minimal immunoglobulins can be designed using

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Recombinant immunoglobulin techniques, MEXICAN INSTITUTE OF PROPERTY. PtS'f, T“'^ Je'i ^ T ¥ o7J Fv immunoglobulins for use herein — inven Cl'STi — can be produced by synthesizing a fused variable light chain region and a variable heavy chain region. Antibody combinations, eg, diabodies, which comprise two distinct Fv specificities are also of interest. In another embodiment of the invention, SMIPs (small molecule immunological pharmaceuticals), camelid antibodies, nano-antibodies and IgNARs are comprised in the immunoglobulin fragments.

Immunoglobulins and fragments thereof can be post-translationally modified, for example, to add effector moieties such as chemical bonds, detectable moieties, such as fluorescent dyes, enzymes, toxins, substrates, bioluminescent materials, radioactive materials, chemiluminescent moieties, and the like, or may use specific binding moieties, such as streptavidin, avidin or biotin and the like in the methods and compositions of the present invention. Examples of additional effector molecules are provided below.

The term polyploid yeast that stably expresses or expresses a desired secreted heterologous polypeptide for a long time refers to a yeast culture that secretes said polypeptide for at least several days to a week, more preferably, at least a month,

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even more preferably, at least 1-6 months and, even more preferably, for more than a year at threshold expression levels, typically at least 10-25 mg / liter and preferably considerably higher.

The term "polyploid yeast culture" that secretes desired amounts of recombinant polypeptide refers to cultures that stably or for long periods of time secrete at least 10-25 mg / liter of heterologous polypeptide, more preferably, at least 50-500 mg / liter and , more preferably, 500-1000 mg / liter or more.

A polynucleotide sequence corresponds to a polypeptide sequence if translation of the polynucleotide sequence according to the genetic code produces the polypeptide sequence (ie, the polynucleotide sequence encodes the polypeptide sequence); one polynucleotide sequence corresponds to another polynucleotide sequence if the two sequences encode the same polypeptide sequence.

A region or heterologous domain of a DNA construct is an identifiable segment of DNA within a larger DNA molecule that is not associated with the largest molecule in nature. Accordingly, when the heterologous region encodes a mammalian gene, the gene will generally be flanked by DNA that does not flank mammalian genomic DNA in the original organism genome. Other

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An example of a heterologous region is a construct in which the coding sequence itself is not found in nature (for example, a cDNA where the genomic coding sequence contains introns or synthetic sequences that possess codons other than native gene). Allelic variations or naturally occurring mutational events do not generate a heterologous region of DNA as defined herein.

A coding sequence is a sequence within the codon framework that (in view of the genetic code) corresponds to or encodes a protein or peptide sequence. Two coding sequences correspond to each other if the sequences or their complementary sequences encode the same amino acid sequences. A coding sequence associated with appropriate regulatory sequences can be transcribed and translated into a polypeptide. A polyadenylation signal and a transcription termination sequence will generally be located 3 'with respect to the coding sequence. A promoter sequence is a regulatory region of DNA that can bind to RNA polymerase in a cell and initiate transcription of a downstream coding sequence (3 'direction). Promoter sequences typically contain additional sites for binding regulatory molecules (eg, transcription factors) that affect transcription of the coding sequence.

A coding sequence is under the control of the promoter sequence or operably linked to the promoter when RNA polymerase binds the promoter sequence in a cell and transcribes the coding sequence into mRNA, which is then translated into the protein encoded by the coding sequence.

Vectors are used to introduce a foreign substance, such as DNA, RNA, or protein into an organism or a host cell. Typical vectors include recombinant viruses (for polynucleotides) and liposomes or other lipid aggregates (for polypeptides and / or polynucleotides). A DNA vector is a replicon, such as a plasmid, phage, or cosmid, to which another polynucleotide segment can be attached to cause replication of the attached segment. An expression vector is a DNA vector that contains regulatory sequences that will direct the synthesis of polypeptides by an appropriate host cell. This in general means a promoter to bind RNA polymerase and initiate mRNA transcription, as well as ribosome binding sites and initiation signals to direct translation of the mRNA into one or more polypeptides. Incorporation of a polynucleotide sequence into an expression vector at the appropriate site and in the correct reading frame, followed by transformation of an appropriate host cell by the vector allows the production of a polypeptide encoded by

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said polynucleotide sequence. Examples of expression vectors and techniques for their use are described in the following publications: Oíd et al., Principies of Gene Manipulation: An Introduction to Genetic Engineering, Blackwell Scientific

Publications, 4to edition, 1989; Sambrook et al., Molecular

Cloning: A Laboratory Manual, 2to edition, Coid Spring Harbor

Laboratory Press, 1989; Sambrook et al., Molecular Cloning: A

Laboratory Manual, 3to Edition, Coid Spring Harbor Laboratory

Press, 2001; Gorman, High Efficiency Gene Transfer into

Mammalian Cells, in DNA Cloning, Volume II, Glover, DM,

Ed., IRL Press, Washington, DC, pp. 143 190 (1985).

For example, a liposome or other lipid aggregate can comprise a lipid, such as phosphatidylcholines (lecithins) (PC), phosphatidylethanolamines (PE), lysolecithins, lysophosphatidylethanolamines, phosphatidylserines (PS), phosphatidylglycerols (PG), phosphatidylinositol (PI), cardiolipin, phosphatidic acids (PA), fatty acids, gangliosides, glycolipids, glycolipids, mono-, di or triglycerides, ceramides, cerebrosides and combinations thereof; a cationic lipid (or other cationic amphiphilic) such as 1,2-dioleyloxy-3- (trimethylamino) propane (DOTAP), Ncolesteriloxycarbaryl-3, 7,12-triazapentadecane-l, 15-diamine (CTAP), N- bromide [1- (2,3, -ditetradecyloxy) propyl] -N, Ndimethyl-N-hydroxyethylammonium (DMRIE), N- [1- (2,3, dioleyloxy) propyl] -N, N-dimethyl-N- bromide hydroxy ethylammonium (DORIE),

IMPIOS

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL Aj ^ -x¿j— N- [1- (2,3-dioleyloxy) propyl] -N, N, N-trimethylammonium chloride (DOTMA), 3 beta [N- (N ', N'-dimethylaminoethane) carbamoyl] cholesterol (DC-Choi) and dimethyldioctadecylammonium (DDAB), dioleoylphosphatidyl ethanolamine (DOPE), cholesterol-containing DOPC and combinations of these and / or a hydrophilic polymer such as polyvinylpyrrolidylarylpyrilatolipheroxylazyl, polymethyloxazoline, polyvinyloxylazaryl, , polydimethylacrylamide, polyhydroxypropylmethacrylate, polyhydroxyethyl acrylate, hydroxymethyl cellulose, hydroxyethyl cellulose, polyethylene glycol, polyaspartamide, and combinations thereof. Other suitable cationic lipids are described in Miller, Angew. Chem. Int. Ed. 37: 1768 1785 (1998), and Cooper et al., Chem. Eur. J. 4 (1):

137 151 (1998). The liposomes can be crosslinked, partially crosslinked, or non-crosslinked. Crosslinked liposomes can include crosslinked as well as non-crosslinked components. Suitable cytofectins or cationic liposomes are commercially available and can also be prepared as described in Sipkins et al., Nature.

Medicine, 1998, 4 (5) :( 1998), 623 626 or described in Miller, supra. Examples of liposomes include a polymerizable neutral or zwitterionic lipid, a polymerizable integrin targeting lipid, and a polymerizable cationic lipid suitable for binding of a nucleic acid. Liposomes can

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL optionally include peptides that provide increased efficacy, for example, as described in US Patent No. 7,297,759. Additionally, examples of liposomes and other lipid aggregates are described in US Patent No.

7,166,298.

Polynucleotide sequence amplification is the in vitro production of multiple copies of a particular nucleic acid sequence. The amplified sequence is generally in the form of DNA. A variety of techniques for carrying out such amplification are described in a review by Van

Brunt (1990, Bio / Technol., 8 (4): 291-294). Polymerase chain reaction or PCR is a prototype nucleic acid amplification and the use of PCR herein should be considered an example of other suitable amplification techniques.

Currently, the general structure of antibodies in vertebrates is well understood (Edelman, G. Μ., Ann. NY Acad.

Sci, 190: 5 (1971)). Antibodies consist of two identical polypeptide light chains with a molecular weight of approximately 23,000 daltons (the light chain) and two identical heavy chains with a molecular weight of 53,00070,000 (the heavy chain). The four chains are joined by disulfide bonds in a Y configuration where the light chains group the heavy chains starting at the mouth of the Y configuration. The branch portion of the

configuration Y is designated as region Ftob; the stem portion of the Y configuration is designated the Fe region. The orientation of the amino acid sequence runs from the extreme

N-terminal at the beginning of the Y configuration to the C-terminal end at the end of each string. The N-terminus has the variable region that has specificity for the antigen that caused it and is approximately 100 amino acids in length. However, there are small variations between light and heavy chain, as well as from antibody to antibody.

The variable region binds in each chain to a constant region that spans the remaining length of the chain and that within a particular class of antibody does not vary with the specificity of the antibody (i.e., the antigen that causes it). There are five main known classes of constant regions that determine the immunoglobulin molecule class (IgG, IgM, IgA, IgD, and IgE that correspond to the heavy chain constant regions γ, μ, α, δ, and ε (gamma, mu, alpha, delta, or epsilon.) The constant region or class determines the subsequent effector function of the antibody, including complement activation (Kabat, EA, Structural Concepts in Immunology and Immunochemistry, 2to Ed., P. 413-436, Holt, Rinehart, Winston (1976)), and other cellular responses (Andrews, DW, et al., Clinical Immunobiology, pp 1-18, WB Sanders (1980); Kohl, S., et al., Immunology, 48: 187 (1983)), while the variable region

determines the antigen with which it will react. Light chains are classified as κ (kappa) or λ (lambda). Each kind of heavy chain can be paired with a kappa or lambda light chain. The light and heavy chains are covalently linked to each other and the tail portions of the two heavy chains are linked to each other by means of covalent disulfide bonds when immunoglobulins are generated by hybridomas or B lymphocytes.

The term "variable region" or "VR" refers to the domains within each pair of light and heavy chains in an antibody that are directly involved in binding the antibody to the antigen. Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end, the constant domain of the light chain is aligned with the first constant domain of the heavy chain and the variable domain of the light chain is aligned with the variable domain of the heavy chain.

The terms "complementarity determining region", "hypervariable region" or "CDR" refer to one or more of the "complementary or hyper variable determining regions" (CDR) that are found in the variable regions of the light or heavy chains of an antibody (See Kabat, EA et al. al., Sequences of Proteins of

IMP

M2XICA INSTITUTE OF THE PROPIEU INDUSTR

Immunological Interest, National Institutes of Health,

Bethesda, Md., (1987)). These expressions include hypervariable regions as defined in Kabat et al. (Sequences of Proteins of Immunological Interest, Kabat E., et al., US

Dept. of Health and Human Services, 1983) or hypervariable loops in three-dimensional antibody structures (Chothia and Lesk, J Mol. Biol. 196 901-917 (1987)). The CDRs of each chain are maintained in close proximity by flanking regions and, with the CDRs of the other chain, contribute to the formation of the antigen binding site. Within the CDRs are select amino acids that have been described as the selectivity determining regions (SDRs) that represent the critical contact residues used by the CDRs in the antibody-antigen interaction (Kashmiri, S., Methods, 36: 2534 (2005 )).

The terms flanking region or FR refer to one or more of the flanking regions within the variable regions of the light or heavy chains of an antibody (See Kabat, EA et al., Sequences of Proteins of Immunological Interest, National Institutes of Health,

Bethesda, Md., (1987)). These expressions include the regions of amino acid sequences interposed between CDRs within the variable regions of the light and heavy chains of an antibody.

IMPI

MEXICAN INSTITUTE OF LA ??. O? L £ üAD industrial

Anti-IL-6 antibodies and binding fragments thereof

The invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKPG

QRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNAFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN (SEQ ID NO: 2) or

AIQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFS

GSGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNAFGGGTKVEIKR (SEQ ID NO:

709) .

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYVTWVRQAPGKGL

EWIGIIYGSDETAYATWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNLW

GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK (SEQ ID NO: 3) or

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATSA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSS (SEQ

ID NO: 657).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 4; SEQ ID NO: 5 and SEQ ID NO: 6 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 2 or 709, and / or one or more of the sequences of

κ hee

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL polypeptides of SEQ ID NO: 7; SEQ ID NO: 8 and SEQ ID NO: 9 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 3 or 657 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 4; SEQ ID NO: 5 and SEQ ID NO: 6 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 2 or 709, and / or one or more of the polypeptide sequences of SEQ ID NO: 7; SEQ ID NO: 8 and SEQ ID NO: 9 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 3 or 657 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In a modality of acó »

I

INSTITUTO MEXICANO DE LA PROPERTY INDUSTRIAL the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 2 or 657. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, a polypeptide sequence of SEQ ID NO: 3 or 709.

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 4; SEQ ID NO: 5 and SEQ ID NO: 6 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 2nd SEQ ID

NO: 709.

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 7; SEQ ID NO: 8 and SEQ ID NO: 9 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 3 and SEQ

ID NO: 657.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, fragments of

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

Antibodies possessing IL-6 binding specificity comprise or alternatively consist of one, two, three or more antibody fragments, including all of the following: the variable light chain region of SEQ ID

NO: 2 or 709; the variable heavy chain region of SEQ ID

NO: 3 or 657; the complementarity determining regions (SEQ ID NO: 4; SEQ ID NO: 5 and SEQ ID NO: 6) of the variable light chain region of SEQ ID NO: 2 or 709; and the complementarity determining regions (SEQ ID NO: 7; SEQ ID NO: 8 and SEQ ID NO: 9) of the variable heavy chain region of the

SEQ ID NO: 3 or SEQ ID NO: 657.

The invention also contemplates variants where any of the heavy chain polypeptide sequences of SEQ ID NO: 18 or SEQ ID NO: 19 replace the heavy chain polypeptide sequence of SEQ ID NO: 3 or 657, the sequence of light chain polypeptide of SEQ ID NO: 20 replaces the light chain polypeptide sequence of

SEQ ID NO: 2 or SEQ ID NO: 709 and the heavy chain CDR sequence of SEQ ID NO: 120 replaces the heavy chain CDR sequence of SEQ ID NO: 8.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl, comprising SEQ ID NO: 2 and SEQ ID NO: 3, or an antibody comprising SEQ ID NO: 657 and SEQ ID NO: 709 (which are respectively encoded by the nucleic acid sequences in SEQ ID NO: 700 and SEQ ID impious

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INDUSTRIAL

NO: 723) one comprised of the alternative SEQ ID NO sequences set forth in the preceding paragraph, or comprised in Figures 34-37 and having at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTINCQASETIYSWLSWYQQKPG

QPPKLLIYQASDLASGVPSRFSGSGAGTEYTLTISGVQCDDAATYYCQQGYSGSNVDNVFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFY (SEQ ID NO: 21)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQEQLKESGGRLVTPGTPLTLTCTASGFSLNDHAMGWVRQAPGKG

LEYIGFINSGGSARYASWAEGRFTISRTSTTVDLKMTSLTTEDTATYFCVRGGAVWSIHSFDP

WGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK (SEQ ID NO: 22).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 23; SEQ ID NO: 24 and SEQ ID NO: 25 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 21 and / or one or more of the polypeptide sequences from SEQ ID NO: 26; SEQ ID NO: 27 and SEQ ID NO: 28 corresponding to the determining regions of complementarity

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY (CDR or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 22 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 23; SEQ ID NO: 24 and SEQ ID NO: 25 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 21 and / or one or more of the polypeptide sequences from SEQ ID NO: 26; SEQ ID NO: 27 and SEQ ID NO: 28 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 22 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

of polypeptides of SEQ ID NO: 21. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, a sequence of polypeptides of SEQ ID NO: 22.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 23; SEQ ID NO:

and SEQ ID NO: 25 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 21.

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or alternatively consist of one or more polypeptide sequences of SEQ ID NO: 26; SEQ ID NO: 27 and SEQ ID NO: 28 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 22.

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INDUSTRIAL

three or more antibody fragments, including all of the following: the variable light chain region of SEQ ID NO: 21, the variable heavy chain region of SEQ ID NO: 22, the complementarity determining regions (SEQ ID NO:

2. 3; SEQ ID NO: 24 and SEQ ID NO: 25) of the variable light chain region of SEQ ID NO: 21 and the complementarity determining regions (SEQ ID NO: 26; SEQ ID NO: 27 and SEQ ID NO: 28 ) of the variable heavy chain region of SEQ ID NO: 22.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab2 comprising SEQ ID NO: 21 and SEQ ID NO: 22, which possesses at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQASQSVYDNNYLSWFQQK

PGQPPKLLIYGASTLASGVPSRFVGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDNAFGG

GTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN (SEQ ID NO: 37)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSVYYMNWVRQAPGKGL

EWIGFITMSDNINYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARSRGWGTMGRLDLW

100

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INDUSTRIAL

GPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK (SEQ ID NO: 38).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 37 and / or one or more of the polypeptide sequences from SEQ ID NO: 42; SEQ ID NO: 43 and SEQ ID NO: 44 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 38 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

101

Variable Heavy INDUSTRIAL PROPERTY MEXICAN INSTITUTE OF SEQ ID NO: 38 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 37. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 38.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 39; SEQ ID NO:

and SEQ ID NO: 41 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 37.

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INSTITUTE ME.'CCAMO DE LA kROf'iSDAD

INDUSTRIAL

and SEQ ID NO: 44 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 38.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, antibody fragments possessing IL-6 binding specificity comprise or alternatively consist of one, two, three or more antibody fragments, including all of the following: the strand region light variable of SEQ ID NO: 37, the variable heavy chain region of SEQ ID NO: 38, the complementarity determining regions (SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41) of the variable light chain region of SEQ ID NO: 37 and the complementarity determining regions (SEQ ID NO: 42; SEQ ID NO:

and SEQ ID NO: 44) of the variable heavy chain region of SEQ ID NO: 38..

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab3 comprising SEQ ID NO: 37 and SEQ ID NO: 38, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies that possess IL-6 binding specificity and a variable light chain sequence that comprises the sequence set out at

103

continuation:

INSTITUTO CT & .NO DE U i'RO / CO.W INDUJTKÍAt

MDTRAPTQLLGLLLLWLPGAICDPVLTQTPSPVSAPVGGTVSISCQASQSVYENNYLSWFQQK

PGQPPKLLIYGASTLDSGVPSRFKGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDDAFGG

GTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN (SEQ ID NO: 53)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQEQLKESGGGLVTPGGTLTLTCTASGFSLNAYYMNWVRQAPGKG

LEWIGFITLNNNVAYANWAKGRFTFSKTSTTVDLKMTSPTPEDTATYFCARSRGWGAMGRLDL

WGHGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK (SEQ ID NO: 54).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 53 and / or one or more of the polypeptide sequences from SEQ ID NO: 58; SEQ ID NO: 59 and SEQ ID NO: 60 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 54 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other

104

I

antibodies such as chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 53 and / or one or more of the polypeptide sequences from SEQ ID NO: 58; SEQ ID NO: 59 and SEQ ID NO: 60 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 54 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 53. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 54.

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or, alternatively, consist of one or

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more polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 53.

and SEQ ID NO: 60 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 54.

NO: 53, the variable heavy chain region of SEQ ID NO: 54, the complementarity determining regions (SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57) of the variable light chain region of SEQ ID NO: 53 and complementarity determining regions (SEQ ID NO: 58; SEQ ID NO:

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Mexicano ~ iNSTr 'JTÜ Mexican DE LA J-RC.pieoaO l »lu'cL'LRiAL

and SEQ ID NO: 60) of the variable heavy chain region of SEQ ID NO: 54. -

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab4 comprising SEQ ID NO: 53 and SEQ ID NO:

54, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSPVSAAVGGTVTINCQASQSVDDNNWLGWYQQK

RGQPPKYLIYSASTLASGVPSRFKGSGSGTQFTLTISDLECDDAATYYCAGGFSGNIFAFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF (SEQ ID NO: 69)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGL

EWIGIIGGFGTTYYATWAKGRFTISKTSTTVDLRITSPTTEDTATYFCARGGPGNGGDIWGQG

TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD (SEQ ID NO: 70).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 71; SEQ ID NO: 72 and SEQ ID NO: 73 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 69 and / or one or more of the polypeptide sequences

107

from SEQ ID NO: 74; SEQ ID NO: 75 and SEQ ID NO: 76 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 70 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 71; SEQ ID NO: 72 and SEQ ID NO: 73 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 69 and / or one or more of the polypeptide sequences from SEQ ID NO: 74; SEQ ID NO: 75 and SEQ ID NO: 76 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 70 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In a form of

108

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1NDUSTKIAL

In the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 69. In another embodiment of the invention, the antibody fragments of the invention comprise, or in a manner Alternatively they consist of a polypeptide sequence of SEQ ID NO: 70.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 71; SEQ ID NO:

and SEQ ID NO: 73 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 69.

and SEQ ID NO: 76 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 70.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, fragments of

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NO: 69, the variable heavy chain region of SEQ ID NO:

70, the complementarity determining regions (SEQ ID NO:

71; SEQ ID NO: 72 and SEQ ID NO: 73) of the variable light chain region of SEQ ID NO: 69 and the complementarity determining regions (SEQ ID NO: 74; SEQ ID NO: 75 and SEQ ID NO: 7 6) from the variable heavy chain region of SEQ ID NO: 70.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab5 comprising SEQ ID NO: 69 and SEQ ID NO: 70, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSVPVGGTVTIKCQSSQSVYNNFLSWYQQKP

GQPPKLLIYQASKLASGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNAFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF (SEQ ID NO: 85)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

110

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MEXICAN INSTITUTE OF PkCT Ií'DAD

'i / .i wrqápgkgl

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGIDra ^ ftM

EWIGIIYAGSGSTWYASWAKGRFTISKTSTTVDLKITSPl'IWrm ICAREA 1WGW) RL

DLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD (SEQ ID NO: 86).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 87; SEQ ID NO: 88 and SEQ ID NO: 89 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 85 and / or one or more of the polypeptide sequences from SEQ ID NO: 90; SEQ ID NO: 91 and SEQ ID NO: 92 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 86 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 87; SEQ ID NO: 88 and SEQ ID NO: 89 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the | variable light chain sequence of SEQ ID NO: 85 and / or one or more of the sequences of polypeptides of SEQ ID NO: 90; SEQ ID NO: 91 and SEQ ID NO: 92 which

111

they correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 86 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 85. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 86.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 87; SEQ ID NO: 88 and SEQ ID NO: 89 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 85.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

112

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INSTITUTO VE'CC.-NO

Say La? XC? ÍtiMD V

IL-6 comprise or alternatively consist of one or more polypeptide sequences of SEQ 'TÚ) NO: W; SEQ ib NO: 91 and SEQ ID NO: 92 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 86.

87; SEQ ID NO: 88 and SEQ ID NO: 89) of the variable light chain region of SEQ ID NO: 85 and the complementarity determining regions (SEQ ID NO: 90; SEQ ID NO:

and SEQ ID NO: 92) of the variable heavy chain region of SEQ ID NO: 86.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab6 comprising SEQ ID NO: 85 and SEQ ID NO:

86, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies that

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INSTITUTO MEXICANO DE LA PROPIEDAD INDUSTRIAL have specificity for binding to IL-6 and a variable light chain sequence that includes the following sequence:

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKSG

QRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNAFGGG

TEWVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF (SEQ ID NO: 101)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLSGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYMTWVRQAPGKGL

EWIGMIYGSDETAYANWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNLW

GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK (SEQ ID NO: 102).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 103; SEQ ID NO: 104 and SEQ ID NO: 105 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 101 and / or one or more of the polypeptide sequences from SEQ ID NO: 106; SEQ ID NO: 107 and SEQ ID NO: 108 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 102 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable light and heavy chain sequences

114

previously established. _

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 103; SEQ ID NO: 104 and SEQ ID NO: 105 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 101 and / or one or more of the polypeptide sequences from SEQ ID NO: 106; SEQ ID NO: 107 and SEQ ID NO: 108 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 102 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 101. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 102.

In a further embodiment of the invention, the

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JL 1L

MEXICAN INSTITUTE>

OF THE IHOPIEDAD \

INDUSTRIAL antibody fragments that possess binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 103; SEQ ID NO:

104 and SEQ ID NO: 105 corresponding to the determining regions of (hypervariable CDR complementarity) of the variable light chain sequence of the

SEQ ID NO: 101.

regions

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 106; SEQ ID NO:

107 and SEQ ID NO: 108 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 102.

102, the complementarity determining regions (SEQ ID NO: 103; SEQ ID NO: 104 and SEQ ID NO: 105) of the region of

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Variable light chain of SEQ ID NO: 101 and the «III Ι-Χ2.ΎAíií-J ... Si-sesv complementary determinants regions (SEQ ID NO: 106; SEQ ID NO: 107 and SEQ ID NO: 108) of the variable heavy chain region of SEQ ID NO: 102.

The invention also contemplates variants where any of the heavy chain polypeptide sequences of SEQ ID

NO: 117 or SEQ ID NO: 118 replace the heavy chain polypeptide sequence of. SEQ ID NO: 102; the light chain polypeptide sequence of SEQ ID NO: 119 replaces the light chain polypeptide sequence of SEQ ID NO: 101 and the heavy chain CDR sequence of SEQ ID NO: 121 replaces the sequence of Heavy chain CDR of SEQ ID NO: 107.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab7 comprising SEQ ID NO: 101 and SEQ ID

NO: 102, or the alternative SEQ ID NO sequences set forth in the preceding paragraph, and having at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQSSQSVGNNQDLSWFQQR

PGQPPKLLIYEISKLESGVPSRFSGSGSGTHFTLTISGVQCDDAATYYCLGGYDDDADNA (SEQ ID NO: 122)

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The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCHSVEESGGRLVTPGTPLTLTCTVSGFSLSSRTMSWVRQAPGKGL

EWIGYIWSGGSTYYATWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARLGDTGGHAYATRL

NL (SEQ ID NO: 123).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 124; SEQ ID NO: 125 and SEQ ID NO: 126 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 122 and / or one or more of the polypeptide sequences from SEQ ID NO: 127; SEQ ID NO: 128 and SEQ ID NO: 129 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 123 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 124; SEQ ID NO: 125 and SEQ ID NO: 126 that correspond to the complementarity determining regions (CDR or regions

118

hypervariables) of the variable light chain sequence of SEQ ID NO: 122 and / or one or more of the polypeptide sequences of SEQ ID NO: 127; SEQ ID NO: 128 and SEQ ID NO: 129 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 123 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 122. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 123.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 124; SEQ ID NO: 125 and SEQ ID NO: 126 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

119

SEQ ID NO: 122.

In a further embodiment of the invention, fragments of the antibody that possess IL-6 binding specificity comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 127; SEQ ID NO:

128 and SEQ ID NO: 129 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 123.

NO: 122, the variable heavy chain region of SEQ ID NO: 123, the complementarity determining regions (SEQ ID NO: 124; SEQ ID NO: 125 and SEQ ID NO: 126) of the variable light chain region of SEQ ID NO: 122 and the complementarity determining regions (SEQ ID NO: 127; SEQ ID NO: 128 and SEQ ID NO: 129) of the variable heavy chain region of SEQ ID NO: 123.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab8 comprising SEQ ID NO: 122 and SEQ ID

120

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NO: 123, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSSVSAAVGGTVSISCQSSQSVYSNKYLAWYQQK

PGQPPKLLIYWTSKLASGAPSRFSGSGSGTQFTLTISGVQCDDAATYYCLGAYDDDADNA (SEQ ID NO: 138)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVKPDETLTLTCTASGFSLEGGYMTWVRQAPGKGL

EWIGISYDSGSTYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCVRSLKYPTVTSDDL (SEQ ID NO: 139).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 140; SEQ ID NO: 141 and SEQ ID NO: 142 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 138 and / or one or more of the polypeptide sequences from SEQ ID NO: 143; SEQ ID NO: 144 and SEQ ID NO: 145 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 139 or combinations thereof

121

polypeptide sequences. In another embodiment of jnvpnpiAn, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 140; SEQ ID NO: 141 and SEQ ID NO: 142 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 138 and / or one or more of the polypeptide sequences of SEQ ID NO: 143; SEQ ID NO: 144 and SEQ ID NO: 145 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 139 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 138. In another embodiment of the invention, the antibody fragments of the invention

122

comprise, or alternatively conist on, a polypeptide sequence of SEQ ID NO: 139.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 140; SEQ ID NO:

141 and SEQ ID NO: 142 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 138.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 143; SEQ ID NO: 144 and SEQ ID NO: 145 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 139.

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INDUSTRIAL

NO: 138, the variable heavy chain region of SEQ ID NO: 139, the complementarity determining regions (SEQ ID NO: 140; SEQ ID NO: 141 and SEQ ID NO: 142) of the variable light chain region of SEQ ID NO: 138 and the complementarity determining regions (SEQ ID NO: 143; SEQ ID NO: 144 and SEQ ID NO: 145) of the variable heavy chain region of SEQ ID NO: 139.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab9 comprising SEQ ID NO: 138 and SEQ ID NO: 139, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQT PS PVSAAVGGTVTISCQS SQSVYNNNDLAWYQQK

PGQPPKLLIYYASTLASGVPSRFKGSGSGTQFTLTISGVQCDDAAAYYCLGGYDDDADNA (SEQ ID NO: 154)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGLSLSSNTINWVRQAPGKGL

EWIGYIWSGGSTYYASWVNGRFTISKTSTTVDLKITSPTTEDTATYFCARGGYASGGYPYATR

LDL (SEQ ID NO: 155).

The invention further contemplates antibodies that

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comprise one or more polypeptide sequences of SEQ ID ———— b · —ι · ιι · ~ * i *

NO: 156; SEQ ID NO: 157 and SEQ ID NO: 158 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 154 and / or one or more of the polypeptide sequences from SEQ ID NO: 159; SEQ ID NO: 160 and SEQ ID NO: 161 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 155 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

125

Institute

To the industrial PYOFIEDaO the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 154. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 155.

157 and SEQ ID NO: 158 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 154.

160 and SEQ ID NO: 161 corresponding to the complementarity determining regions (CDR or regions

126

GIVE THE PROPERTY hypervariables) of the heavy chain sequence1'variable'

SEQ ID NO: 155.

NO: 154, the variable heavy chain region of SEQ ID NO:

155, the complementarity determining regions (SEQ ID NO: 156; SEQ ID NO: 157 and SEQ ID NO: 158) of the variable light chain region of SEQ ID NO: 154 and the complementarity determining regions (SEQ ID NO: 154). : 159; SEQ ID NO:

160 and SEQ ID NO: 161) of the variable heavy chain region of SEQ ID NO: 155.

In a preferred embodiment of the invention, the anti-IL-6 antibody is AblO comprising SEQ ID NO: 154 and SEQ ID

NO: 155, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSSVSAAVGGTVTINCQSSQSVYNNDYLSWYQQR

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MEXICAN INSTITUTE nc i A 9νηΡΐΠ) ΑΠ

PGQRPKLLIYGASKLASGVPSRFKGSGSGKQFTLTISGVQCDDAATYYCLGDYDDDADNT (SEQ ID NO: 170)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFTLSTNYYLSWVRQAPGKG

LEWIGIIYPSGNTYCAKWAKGRFTISKTSSTTVDLKMTSPTTEDTATYFCARNYGGDESL (SEQ ID NO: 171).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 172; SEQ ID NO: 173 and SEQ ID NO: 174 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 170 and / or one or more of the polypeptide sequences from SEQ ID NO: 175; SEQ ID NO: 176 and SEQ ID NO: 177 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 171 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

128

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence I of SEQ ID NO: 170. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of a polypeptide sequence of SEQ ID NO: 171.

129 *

INSTITUTO MEXICzINO D6 LA PROPIIOAD INDUSTRIAL complementary determinants (CDR or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 170.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 175; SEQ ID NO: 176 and SEQ ID NO: 177 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 171.

NO: 170, the variable heavy chain region of SEQ ID NO: 171, the complementarity determining regions (SEQ ID

NO: 172; SEQ ID NO: 173 and SEQ ID NO: 174) of the variable light chain region of SEQ ID NO: 170 and the complementarity determining regions (SEQ ID NO: 175; SEQ ID NO: 176 and SEQ ID NO: 177 ) of the variable heavy chain region of SEQ ID NO: 171.

130

In a preferred embodiment of the

MEXICAN INSTITUTE D £ THE PROPERTY

INDUSTRIAL invention, the

anti-IL-6 antibody is Abll comprising SEQ ID NO: 170 and SEQ ID NO: 171, which possesses at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASETIGNALAWYQQKSG

QPPKLLIYKASKLASGVPSRFKGSGSGTEYTLTISDLECADAATYYCQWCYFGDSV (SEQ

ID NO: 186)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVTVLKGVQCQEQLVESGGGLVQPEGSLTLTCTASGFDFSSGYYMCWVRQAPGK

GLEWIACIFTITTNTYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYLCARGIYSDNNYY

AL (SEQ ID NO: 187).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 188; SEQ ID NO: 189 and SEQ ID NO: 190 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 186 and / or one or more of the polypeptide sequences from SEQ ID NO: 191; SEQ ID NO: 192 and SEQ ID NO: 193 corresponding to the complementarity determining regions

131

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY (CDR or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 187 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 188; SEQ ID NO: 189 and SEQ ID NO: 190 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 186 and / or one or more of the polypeptide sequences from SEQ ID NO: 191; SEQ ID NO: 192 and SEQ ID NO: 193 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 187 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In an embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the sequence

132

of polypeptides of SEQ ID NO: 18 6. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, a sequence of polypeptides of SEQ ID NO: 187.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 188; SEQ ID NO:

189 and SEQ ID NO: 190 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 186.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 191; SEQ ID NO:

192 and SEQ ID NO: 193 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 187.

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MEXICAN INSTITUTE OF THE ERCPIED. · D industrial

three or more antibody fragments, including all of the following: the variable light chain region of ^ St¡0 ID 'NO: 186, the variable heavy chain region of SEQ ID NO:

187, the complementarity determining regions (SEQ ID

NO: 188; SEQ ID NO: 189 and SEQ ID NO: 190) of the variable light chain region of SEQ ID NO: 186 and the complementarity determining regions (SEQ ID NO: 191; SEQ ID NO: 192 and SEQ ID NO: 193 ) of the variable heavy chain region of SEQ ID NO: 187.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl2 comprising SEQ ID NO: 186 and SEQ ID NO: 187, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASESIGNALAWYQQKPG

QPPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISGVQCADAAAYYCQWCYFGDSV (SEQ

ID NO: 202)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQQQLVESGGGLVKPGASLTLTCKASGFSFSSGYYMCWVRQAPGK

GLESIACIFTITDNTYYANWAKGRFTISKPSSPTVTLQMTSLTAADTATYFCARGIYSTDNYY

134

AL (SEQ ID NO: 203).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 204; SEQ ID NO: 205 and SEQ ID NO: 206 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 202 and / or one or more of the polypeptide sequences of SEQ ID NO: 207; SEQ-ID NO: 208 and SEQ ID NO: 209 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 203 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 204; SEQ ID NO: 205 and SEQ ID NO: 206 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 202 and / or one or more of the polypeptide sequences of SEQ ID NO: 207; SEQ ID NO: 208 and SEQ ID NO: 209 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the chain sequence

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variable weighing of SEQ ID NO: 203 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 202. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 203.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 204; SEQ ID NO: 205 and SEQ ID NO: 206 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 202.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 207; SEQ ID NO:

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208 and SEQ ID NO: 209 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 203.

NO: 202, the variable heavy chain region of SEQ ID NO: 203, the complementarity determining regions (SEQ ID NO: 204; SEQ ID NO: 205 and SEQ ID NO: 206) of the variable light chain region of SEQ ID NO: 202 and complementarity determining regions (SEQ ID NO: 207; SEQ ID NO:

208 and SEQ ID NO: 209) of the variable heavy chain region of SEQ ID NO: 203.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl3 comprising SEQ ID NO: 202 and SEQ ID

NO: 203, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies that possess IL-6 binding specificity and a variable light chain sequence that comprises the sequence set out at

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continuation:

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASQSVSSYLNWYQQKPG

QPPKLLIYRASTLESGVPSRFKGSGSGTEFTLTISDLECADAATYYCQCTYGTSSSYGAA (SEQ ID NO: 218)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGISLSSNAISWVRQAPGKGL

EWIGIISYSGTTYYASWAKGRFTISKTSSTTVDLKITSPTTEDTATYFCARDDPTTVMVMLIP

FGAGMDL (SEQ ID NO: 219).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 220; SEQ ID NO: 221 and SEQ ID NO: 222 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 218 and / or one or more of the polypeptide sequences from SEQ ID NO: 223; SEQ ID NO: 224 and SEQ ID NO: 225 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 219 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other

138

INSTITUTO MEXICANO DE LA PkOP.'P.OaI) antibodies such as, for example, antibodies qui'ffl.'é ^ ífcosji ^ —which comprise one or more sequences of polypeptides — c3F=ta 'OTSE © “” TB NO: 220; SEQ ID NO: 221 and SEQ ID NO: 222 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 218 and / or one or more of the polypeptide sequences of SEQ ID NO: 223; SEQ ID NO: 224 and SEQ ID NO: 225 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 219 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 218. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 219.

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or, alternatively, consist of one or

139

SEQ ID NO:

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY plus polypeptide sequences of SEQ ID NO: 220;

221 and SEQ ID NO: 222 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 218.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 223; SEQ ID NO:

224 and SEQ ID NO: 225 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 219.

NO: 218, the variable heavy chain region of SEQ ID NO: 219, the complementarity determining regions (SEQ ID NO: 220; SEQ ID NO: 221 and SEQ ID NO: 222) of the variable light chain region of SEQ ID NO: 218 and the complementarity determining regions (SEQ ID NO: 223; SEQ ID NO:

140

224 and SEQ ID the SEQ ID NO my former institute

OF THE ΡΠΟΗμΙΛΟ l?

INDUSTRIAL ^ ¼ ...

NO: 225) of the variable heavy chain region of

219.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl4 comprising SEQ ID NO: 218 and SEQ ID

NO: 219, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAQVLTQTASPVSAAVGGTVTINCQASQSVYKNNYLSWYQQK

PGQPPKGLIYSASTLDSGVPLRFSGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSSGDCYA (SEQ ID NO: 234)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPEGSLTLTCTASGFSFSSYWMCWVRQAPGKGL

EWIACIVTGNGNTYYANWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCAKAYDL (SEQ

ID NO: 235).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 236; SEQ ID NO: 237 and SEQ ID NO: 238 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 234 and / or one or more of the polypeptide sequences

141

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY of SEQ ID NO: 239; SEQ ID NO: 240 and SEQ ID NO: 241 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 235 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 236; SEQ ID NO: 237 and SEQ ID NO: 238 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 234 and / or one or more of the polypeptide sequences from SEQ ID NO: 239; SEQ ID NO: 240 and SEQ ID NO: 241 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 235 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In a form of

142 á JL jí. , _

MEXICAN INSTITUTE 'Tt * / Í

DS LA PRUPiEUAD the invention, the antibody fragments of INthe:’HAinvétitrím comprise, or alternatively consisferT '^ iT ^ Ta ”' secilencict 'of polypeptides of SEQ ID NO: 234. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, a polypeptide sequence of SEQ ID NO: 235.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 236; SEQ ID NO: 237 and SEQ ID NO: 238 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 234.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 239; SEQ ID NO:

240 and SEQ ID NO: 241 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 235.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, fragments of

143

IMPI

Antibodies possessing IL-6 binding specificity Mr. XICAN INSTITUTE OF INDUSTRIAL PROPERTY comprise or alternatively consist of one, two, three or more antibody fragments, including all of the following: the variable light chain region of the SEQ ID

NO: 234, the variable heavy chain region of SEQ ID NO:

235, the complementarity determining regions (SEQ ID

NO: 236; SEQ ID NO: 237 and SEQ ID NO: 238) of the variable light chain region of SEQ ID NO: 234 and the complementarity determining regions (SEQ ID NO: 239; SEQ ID NO:

240 and SEQ ID NO: 241) of the variable heavy chain region of SEQ ID NO: 235.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl5 comprising SEQ ID NO: 234 and SEQ ID NO: 235, which possesses at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGSTFAAVLTQTPSPVSAAVGGTVSISCQASQSVYDNNYLSWYQQK

PGQPPKLLIYGASTLASGVPSRFKGTGSGTQFTLTITDVQCDDAATYYCAGVFNDDSDDA (SEQ ID NO: 250)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

144

METGLRWLLLVAVPKGVQCQSLEESGGRLVTPGTPLTLTCTLSGFSLSAYYMSWVRQAPGKGL

EWIGFITLSDHISYARWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARSRGWGAMGRLDL (SEQ ID NO: 251).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 252; SEQ ID NO: 253 and SEQ ID NO: 254 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 250 and / or one or more of the polypeptide sequences from SEQ ID NO: 255; SEQ ID NO: 256 and SEQ ID NO: 257 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 251 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, which comprise one or more polypeptide sequences of SEQ ID

145

MEXICAN INSTITUTE OF LA PROPISOAl)

INDUSTRIAL

they correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 251 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 250. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 251.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 252; SEQ ID NO: 253 and SEQ ID NO: 254 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 250.

In a fragment of the additional embodiment of the invention, antibodies that possess binding specificity for

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 255; SEQ ID NO: 256 and SEQ ID NO: 257 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 251.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, fragments of antibodies that possess IL-6 binding specificity comprise or alternatively consist of one, two, three or more antibody fragments, including all of the following: the region of variable light chain of SEQ ID NO: 250, the variable heavy chain region of SEQ ID NO:

251, the complementarity determining regions (SEQ ID

NO: 252; SEQ ID NO: 253 and SEQ ID NO: 254) of the variable light chain region of SEQ ID NO: 250 and the complementarity determining regions (SEQ ID NO: 255; SEQ ID NO:

256 and SEQ ID NO: 257) of the variable heavy chain region of SEQ ID NO: 251.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl6 comprising SEQ ID NO: 250 and SEQ ID NO: 251, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies that

147

IMPI

MEXICAN INSTITUTE OF THE INDUSTRIAL PROl'iEOAD

they possess specificity of binding to IL-6 and a variable light chain sequence that comprises the sequence established below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQASQSVYNNKNLAWYQQK

SGQPPKLLIYWASTLASGVSSRFSGSGSGTQFTLTVSGVQCDDAATYYCLGVFDDDADNA (SEQ ID NO: 266)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTASGFSLSSYSMTWVRQAPGKGL

EYIGVIGTSGSTYYATWAKGRFTISRTSTTVALKITSPTTEDTATYFCVRSLSSITFL (SEQ ID NO: 267).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 268; SEQ ID NO: 269 and SEQ ID NO: 270 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 266 and / or one or more of the polypeptide sequences from SEQ ID NO: 271; SEQ ID NO: 272 and SEQ ID NO: 273 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 267 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable light and heavy chain sequences

148

MEXICAN INSTITUTE; ii MfVDIFnAn UV--

previously established.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 268; SEQ ID NO: 269 and SEQ ID NO: 270 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 266 and / or one or more of the polypeptide sequences from SEQ ID NO: 271; SEQ ID NO: 272 and SEQ ID NO: 273 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 267 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 266. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 267.

In a further embodiment of the invention, the

149 - \ 'XV

MEXICAN INSTITUTE J, 1

PROPERTY V ^ -a ^ Sv- -; > INDUSTRIAL fragments of the antibody possessing IL-6 binding specificity comprise or alternatively consist of one or more polypeptide sequences of SEQ ID NO: 268; SEQ ID NO:

269 and SEQ ID NO: 270 which correspond to the 5 complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 266.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 271; SEQ ID NO: 272 and SEQ ID NO: 273 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 267.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, fragments of the antibodies that possess IL-6 binding specificity 20 comprise or, alternatively, consist of one, two, three or more antibody fragments, including all of the following: the region of SEQ ID variable light chain

NO: 266, the variable heavy chain region of SEQ ID NO: 267, the complementarity determining regions (SEQ ID

NO: 268; SEQ ID NO: 269 and SEQ ID NO: 270) from the region of

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-Ut. Λ. -. '. i.

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

variable light chain of SEQ ID NO: 266 and complementarity determining regions (SEQ ID NO: 271; SEQ ID NO:

272 and SEQ ID NO: 273) of the variable heavy chain region of SEQ ID NO: 267.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl7 comprising SEQ ID NO: 266 and SEQ ID

NO: 267, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAFELTQTPASVEAAVGGTVTINCQASQNIYRYLAWYQQKPG

QPPKFLIYLASTLASGVPSRFKGSGSGTEFTLTISDLECADAATYYCQSYYSSNSVA (SEQ

ID NO: 282)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQEQLVESGGDLVQPEGSLTLTCTASELDFSSGYWICWVRQVPGK

GLEWIGCIYTGSSGSTFYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARGYSGFGYF

KL (SEQ ID NO: 283).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 284; SEQ ID NO: 285 and SEQ ID NO: 286 corresponding to the complementarity determining regions (CDRs or regions

151

ρτ

INSTITUTO MEXICANO Lt LA PaONEl'AO industrial hypervariables) of the variable light chain sequence of SEQ ID NO: 282 and / or one or more of the polypeptide sequences of SEQ ID NO: 287; SEQ ID NO: 288 and SEQ ID NO: 289 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 283 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 284; SEQ ID NO: 285 and SEQ ID NO: 286 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 282 and / or one or more of the polypeptide sequences from SEQ ID NO: 287; SEQ ID NO: 288 and SEQ ID NO: 289 which correspond to the complementary complementarity determining regions 20 (CDR or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 283 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence 5 of SEQ ID NO: 282. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, a polypeptide sequence of SEQ ID NO: 283.

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 284; SEQ ID NO: 285 and SEQ ID NO: 286 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 282.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or 20 more polypeptide sequences of SEQ ID NO: 287; SEQ ID NO:

288 and SEQ ID NO: 289 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 283.

The invention also contemplates antibody fragments.

153

INDUSTRIAL including one or more antibody fragments described in

Present. In one embodiment of the invention, fragments 35 "antibodies that possess IL-6 binding specificity comprise or alternatively consist of one, two, three, or more antibody fragments, including all of the following: the region of SEQ ID variable light chain

NO: 282, the variable heavy chain region of SEQ ID NO:

283, the complementarity determining regions (SEQ ID

NO: 284; SEQ ID NO: 285 and SEQ ID NO: 286) of the variable light chain region of SEQ ID NO: 282 and the complementarity determining regions (SEQ ID NO: 287; SEQ ID NO:

288 and SEQ ID NO: 289) of the variable heavy chain region of SEQ ID NO: 283.

In a preferred embodiment of the invention, anti-IL-6 antibody 15 is Abl8 comprising SEQ ID NO: 282 and SEQ ID

NO: 283, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQASEDIYRLLAWYQQKPG

QPPKLLIYDSSDLASGVPSRFKGSGSGTEFTLAISGVQCDDAATYYCQQAWSYSDIDNA (SEQ ID NO: 298)

The invention also includes antibodies that possess

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTASGFSLSSYYMSWVRQAPGKGL

EWIGIITTSGNTFYASWAKGRLTISRTSTTVDLKITSPTTEDTATYFCARTSDIFYYRNL (SEQ ID NO: 299).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 300; SEQ ID NO: 301 and SEQ ID NO: 302 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 298 and / or one or more of the polypeptide sequences from SEQ ID NO: 303; SEQ ID NO: 304 and SEQ ID NO: 305 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 299 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 300; SEQ ID NO: 301 and SEQ ID NO: 302 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

155

MEXICAN INSTITUTE

OF PROPERTY O xstt7 ^ ¿úL¿3r »INDUSTRIAL

SEQ ID NO: 298 and / or one or more of the polypeptide sequences of SEQ ID NO: 303; SEQ ID NO: 304 and SEQ ID NO: 305 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 299 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 298. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 299.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 300; SEQ ID NO: 301 and SEQ ID NO: 302 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 298.

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M £ X INSTITUTE

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or alternatively consist of one or more polypeptide sequences of SEQ ID NO: 303; SEQ ID NO: 304 and SEQ ID NO: 305 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 299.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, antibody fragments possessing IL-6 binding specificity comprise or alternatively consist of one, two, three or more antibody fragments, including all of the following: the strand region light variable of SEQ ID NO: 298, the variable heavy chain region of SEQ ID NO: 299, the complementarity determining regions (SEQ ID NO: 300; SEQ ID NO: 301 and SEQ ID NO: 302) of the variable light chain region of SEQ ID NO: 298 and the complementarity determining regions (SEQ ID NO: 303; SEQ ID NO: 304 and SEQ ID NO: 305 ) of the variable heavy chain region of SEQ ID NO: 299.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Abl9 comprising SEQ ID NO: 298 and SEQ ID

NO: 299, which has at least one of the biological activities

157

MEXICAN INSTITUTE - OF THE PRO? Í¿OAD V ^ ccINDUSTRIAL established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTASPVSAAVGATVTINCQSSQSVYNDMDLAWFQQK

PGQPPKLLIYSASTLASGVPSRFSGSGSGTEFTLTISGVQCDDAATYYCLGAFDDDADNT (SEQ ID NO: 314)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLTRHAITWVRQAPGKGL

EWIGCIWSGGSTYYATWAKGRFTISKTSTTVDLRITSPTTEDTATYFCARVIGDTAGYAYFTG

LDL (SEQ ID NO: 315).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 316; SEQ ID NO: 317 and SEQ ID NO: 318 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 314 and / or one or more of the sequences of polypeptides of SEQ ID NO: 319; SEQ ID NO: 320 and SEQ ID NO: 321 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 315 or combinations of these 25 polypeptide sequences. In another embodiment of the invention,

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IMPI

THE MEXICAN INSTITUTE OF THE PLCPIEOAO INL'USTSlíAL the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 316; SEQ ID NO: 317 and SEQ ID NO: 318 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 314 and / or one or more of the polypeptide sequences from SEQ ID NO: 319; SEQ ID NO: 320 and SEQ ID NO: 321 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 315 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 314. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a sequence

159

I

MEXICAN INSTITUTE L> £ THE PROPERTY

INDUSTRIAL

of polypeptides of SEQ ID NO: 315.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 316; SEQ ID NO:

317 and SEQ ID NO: 318 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 314.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 319; SEQ ID NO: 320 and SEQ ID NO: 321 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 315.

NO: 314, the variable heavy chain region of SEQ ID NO:

160

I

315, the complementarity determining regions (SEQ ID

NO: 316; SEQ ID NO: 317 and SEQ ID NO: 318) of the variable light chain region of SEQ ID NO: 314 and the complementarity determining regions (SEQ ID NO: 319; SEQ ID NO:

320 and SEQ ID NO: 321) of the variable heavy chain region of SEQ ID NO: 315.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab20 comprising SEQ ID NO: 314 and SEQ ID

NO: 315, which possesses at least one of the 10 biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQASQSVYNWLSWYQQKPG

QPPKLLIYTASSLASGVPSRFSGSGSGTEFTLTISGVECADAATYYCQQGYTSDVDNV (SEQ

ID NO: 330)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEEAGGRLVTPGTPLTLTCTVSGIDLSSYAMGWVRQAPGKGL

EYIGIISSSGSTYYATWAKGRFTISQASSTTVDLKITSPTTEDSATYFCARGGAGSGGVWLLD

GFDP (SEQ ID NO: 331).

The invention further contemplates antibodies that comprise one or more polypeptide sequences of SEQ ID.

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MEXICAN INSTITUTE 'C.-T' ^ TS ^ T'ií

OF THE PROPERTY * Λ —---- '-T-Í S

INDUSTRIAL

NO: 332; SEQ ID NO: 333 and SEQ ID NO: 334 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 330 and / or one or more of the polypeptide sequences from SEQ ID NO: 335; SEQ ID NO: 336 and SEQ ID NO: 337 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 331 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 332; SEQ ID NO: 333 and SEQ ID NO: 334 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 330 and / or one or more of the polypeptide sequences from SEQ ID NO: 335; SEQ ID NO: 336 and SEQ ID NO: 337 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 331 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

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INDUS Í R1AL

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 330. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 331.

333 and SEQ ID NO: 334 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 330.

In a further embodiment of the invention, fragments of the antibody possessing specificity for binding to IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 335; SEQ ID NO: 336 and SEQ ID NO: 337 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

SEQ ID NO: 331.

NO: 330, the variable heavy chain region of SEQ ID NO:

331, the complementarity determining regions (SEQ ID NO: 332; SEQ ID NO: 333 and SEQ ID NO: 334) of the variable light chain region of SEQ ID NO: 330 and the complementarity determining regions (SEQ ID NO: 330). : 335; SEQ ID NO: 336 and SEQ ID NO: 337) of the variable heavy chain region of SEQ ID NO: 331.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab21 comprising SEQ ID NO: 330 and SEQ ID NO: 331, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGAKCADVVMTQTPASVSAAVGGTVTINCQASENIYNWLAWYQQKP

GQPPKLLIYTVGDLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSSSYVDNV

164 (SEQ ID NO: 346)

The invention also includes antibodies that possess IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQEQLKESGGRLVTPGTPLTLTCTVSGFSLNDYAVGWFRQAPGKG

LEWIGYIRSSGTTAYATWAKGRFTISATSTTVDLKITSPTTEDTATYFCARGGAGSSGVWILD

GFAP (SEQ ID NO: 347).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 348; SEQ ID NO: 349 and SEQ ID NO: 350 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 346 and / or one or more of the polypeptide sequences from SEQ ID NO: 351; SEQ ID NO: 352 and SEQ ID NO: 353 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 347 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 348; SEQ ID NO: 349 and SEQ ID NO: 350 corresponding to the

165

CIE INDUSTrUAL PROPERTY Complementarity determining regions (CDRs or hypervariables) of the variable light chain sequence of SEQ ID NO: 346 and / or one or more of the polypeptide sequences of SEQ ID NO: 351; SEQ ID NO: 352 and SEQ ID NO: 353 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 347 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 346. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 347.

349 and SEQ ID NO: 350 corresponding to the complementarity determining regions (CDR or regions

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INDUSTRIAL

hypervariables) of the variable light chain sequence

SEQ ID NO: 346.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 351; SEQ ID NO:

352 and SEQ ID NO: 353 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 347.

NO: 346, the variable heavy chain region of SEQ ID NO:

347, the complementarity determining regions (SEQ ID NO: 348; SEQ ID NO: 349 and SEQ ID NO: 350) of the variable light chain region of SEQ ID NO: 346 and the complementarity determining regions (SEQ ID NO: : 351; SEQ ID NO: 352 and SEQ ID NO: 353) of the variable heavy chain region of SEQ ID NO: 347.

In a preferred embodiment of the invention, the antibody

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DE LA rXOrlEü .'- D, INDUSTRIAL anti-IL-6 is Ab22 comprising SEQ ID NO: 346 and NO: 347, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSSVSAAVGGTVTINCQASQSVYQNNYLSWFQQK

PGQPPKLLIYGAATLASGVPSRFKGSGSGTQFTLTISDLECDDAATYYCAGAYRDVDS (SEQ

ID NO: 362)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPGASLTLTCTASGFSFTSTYYIYWVRQAPGKG

LEWIACIDAGSSGSTYYATWVNGRFTISKTSSTTVTLQMTSLTAADTATYFCAKWDYGGNVGW

GYDL (SEQ ID NO: 363).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 364; SEQ ID NO: 365 and SEQ ID NO: 366 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 362 and / or one or more of the polypeptide sequences from SEQ ID NO: 367; SEQ ID NO: 368 and SEQ ID NO: 369 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the chain sequence

168

OF THE VARIABLE HEAVY PKOPÍEDAD OF SEQ ID NO: 363 or combinations do-these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 364; SEQ ID NO: 365 and SEQ ID NO: 366 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 362 and / or one or more of the polypeptide sequences from SEQ ID NO: 367; SEQ ID NO: 368 and SEQ ID NO: 369 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 363 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

169

I saw my Mexican institute.

OF THE PROPERTY

INDUSTRIAL g Invention, the antibody fragments of the invention comprise, or alternatively consist of, a sequence of polypeptides of SEQ ID NO: 363.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 364; SEQ ID NO: 365 and SEQ ID NO: 366 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 362.

368 and SEQ ID NO: 369 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 363.

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OLLA MEXICAN INSTITUTE ΚΚΟίΊΕΟΑΟ

INOUS '.' ÍUAL _ following: the variable light chain region of SEQ ID

NO: 362, the variable heavy chain region of SEQ ID NO:

363, the complementarity determining regions (SEQ ID

NO: 364; SEQ ID NO: 365 and SEQ ID NO: 366) of the variable light chain region of SEQ ID NO: 362 and the complementarity determining regions (SEQ ID NO: 367; SEQ ID NO:

368 and SEQ ID NO: 369) of the variable heavy chain region of SEQ ID NO: 363.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab23 comprising SEQ ID NO: 362 and SEQ ID NO: 363, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAFELTQTPSSVEAAVGGTVTIKCQASQSISSYLAWYQQKPG

QPPKFLIYRASTLASGVPSRFKGSGSGTEFTLTISDLECADAATYYCQSYYDSVSNP (SEQ

ID NO: 378)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPEGSLTLTCKASGLDLGTYWFMCWVRQAPGKG

LEWIACIYTGSSGSTFYASWVNGRFTISKTSSTTVTLQMTSLTAADTATYFCARGYSGYGYFK

L (SEQ ID NO: 379).

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The invention further contemplates antibodies that comprise one or more polypeptide sequences of Ta SEQ'TE NO: 380; SEQ ID NO: 381 and SEQ ID NO: 382 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 378 and / or one or more of the polypeptide sequences of SEQ ID NO: 383; SEQ ID NO: 384 and SEQ ID NO: 385 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 379 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 380; SEQ ID NO: 381 and SEQ ID NO: 382 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 378 and / or one or more of the polypeptide sequences from SEQ ID NO: 383; SEQ ID NO: 384 and SEQ ID NO: 385 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 379 or combinations thereof

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T

polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 378. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 379.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 380; SEQ ID NO: 381 and SEQ ID NO: 382 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 378.

In a further embodiment of the invention, fragments of the antibody possessing specificity for binding to IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 383; SEQ ID NO: 384 and SEQ ID NO: 385 corresponding to the regions

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INSTITUTO MEXICANO DE LA FROPILDAO industrial (CDR hypervariable determinants) of complementarity regions of the variable heavy chain sequence of the

SEQ ID NO: 379.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, antibody fragments possessing IL-6 binding specificity comprise or alternatively consist of one, two, three or more antibody fragments, including all of the following: the strand region light variable of SEQ ID NO: 378, the variable heavy chain region of SEQ ID NO: 379, the complementarity determining regions (SEQ ID NO: 380; SEQ ID NO: 381 and SEQ ID NO: 382) of the variable light chain region of SEQ ID NO: 378 and the complementarity determining regions (SEQ ID NO: 383; SEQ ID NO:

384 and SEQ ID NO: 385) of the variable heavy chain region of SEQ ID NO: 379.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab24 comprising SEQ ID NO: 378 and SEQ ID NO: 379, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

174

MDTRAPTQLLGLLLLWLPGVTFAIEMTQSPFSVSAAVGGTVSISCQASQSVYKNNQLSWYQQK

SGQPPKLLIYGASALASGVPSRFKGSGSGTEFTLTISDVQCDDAATYYCAGAITGSIDTDG (SEQ ID NO: 394)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPGASLTLTCTTSGFSFSSSYFICWVRQAPGKG

LEWIACIYGGDGSTYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCAREWAYSQGYFG

AFDL (SEQ ID NO: 395).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 396; SEQ ID NO: 397 and SEQ ID NO: 398 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 394 and / or one or more of the polypeptide sequences from SEQ ID NO: 399; SEQ ID NO: 400 and SEQ ID NO: 401 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 395 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, which

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comprise one or more polypeptide sequencesriuthe SEQ ID

NO: 396; SEQ ID NO: 397 and SEQ ID NO: 398 corresponding ¿T'Tás Complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 394 and / or one or more of the sequences from polypeptides of SEQ ID NO: 399; SEQ ID NO: 400 and SEQ ID NO: 401 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 395 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 394. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 395.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 396; SEQ ID NO:

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

397 and SEQ ID NO: 398 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 394.

400 and SEQ ID NO: 401 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 395.

NO: 394, the variable heavy chain region of SEQ ID NO:

395, the complementarity determining regions (SEQ ID NO: 396; SEQ ID NO: 397 and SEQ ID NO: 398) of the variable light chain region of SEQ ID NO: 394 and the complementarity determining regions (SEQ ID NO: : 399; SEQ ID NO: 400 and SEQ ID NO: 401) of the variable heavy chain region of

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY SEQ ID NO: 395.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab25 comprising SEQ ID NO: 394 and SEQ ID NO: 395, which possesses at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASEDISSYLAWYQQKPG

QPPKLLIYAASNLESGVSSRFKGSGSGTEYTLTISDLECADAATYYCQCTYGTISISDGNA (SEQ ID NO: 410)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMTWVRQAPGEGL

EYIGFINPGGSAYYASWVKGRFTISKSSTTVDLKITSPTTEDTATYFCARVLIVSYGAFTI (SEQ ID NO: 411).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 412; SEQ ID NO: 413 and SEQ ID NO: 414 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 410 and / or one or more of the polypeptide sequences from SEQ ID NO: 415; SEQ ID NO: 416 and SEQ ID NO: 417 which

178

they correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 411 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 412; SEQ ID NO: 413 and SEQ ID NO: 414 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 410 and / or one or more of the polypeptide sequences from SEQ ID NO: 415; SEQ ID NO: 416 and SEQ ID NO: 417 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 411 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention

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comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 410. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of a polypeptide sequence of SEQ ID NO: 411.

In a further embodiment of the invention, the antibody fragments possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 412; SEQ ID NO:

413 and SEQ ID NO: 414 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 410.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 415; SEQ ID NO:

416 and SEQ ID NO: 417 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 411.

180

The INDUSTRIAL PROPERTY MEXICAN TUTUTU comprise or, alternatively, consist of one, two, three, or more antibody fragments, including all of the following: the variable light chain region of SEQ ID

NO: 410, the variable heavy chain region of SEQ ID NO:

411, the complementarity determining regions (SEQ ID

NO: 412; SEQ ID NO: 413 and SEQ ID NO: 414) of the variable light chain region of SEQ ID NO: 410 and the complementarity determining regions (SEQ ID NO: 415; SEQ ID NO: 416 and SEQ ID NO: 417 ) of the variable heavy chain region of SEQ ID NO: 411.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab26 comprising SEQ ID NO: 410 and SEQ ID NO: 411, which possesses at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWL PGARC DVVMTQT PASVSAAVGGTVTIKCQASE DIE SYLAWYQQKPG

QPPKLLIYGASNLESGVSSRFKGSGSGTEFTLTISDLECADAATYYCQCTYGIISISDGNA (SEQ ID NO: 426)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMTWVRQAPGEGL

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EYIGFMNTGDNAYYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARVLVVAYGAFNI (SEQ ID NO: 427).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 428; SEQ ID NO: 429 and SEQ ID NO: 430 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 426 and / or one or more of the polypeptide sequences from SEQ ID NO: 431; SEQ ID NO: 432 and SEQ ID NO: 433 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 427 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 428; SEQ ID NO: 429 and SEQ ID NO: 430 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 426 and / or one or more of the polypeptide sequences from SEQ ID NO: 431; SEQ ID NO: 432 and SEQ ID NO: 433 corresponding to the determining regions of complementarity

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(CDR or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 427 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 42 6. In another embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, a polypeptide sequence of SEQ ID NO: 427.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 428; SEQ ID NO:

429 and SEQ ID NO: 430 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 426.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or

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more polypeptide sequences of SEQ ID NO: 431; SEQ ID NO: 432 and SEQ ID NO: 433 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 427.

NO: 428; SEQ ID NO: 429 and SEQ ID NO: 430) of the variable light chain region of SEQ ID NO: 426 and the complementarity determining regions (SEQ ID NO: 431; SEQ ID NO:

432 and SEQ ID NO: 433) of the variable heavy chain region of SEQ ID NO: 427.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab27 comprising SEQ ID NO: 426 and SEQ ID NO: 427, which possesses at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies that possess IL-6 binding specificity and a chain sequence

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY light variable comprising the sequence established below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSEPVGGTVSISCQSSKSVMNNNYLAWYQQK

PGQPPKLLIYGASNLASGVPSRFSGSGSGTQFTLTISDVQCDDAATYYCQGGYTGYSDHGT (SEQ ID NO: 442)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVKPDETLTLTCTVSGIDLSSYPMNWVRQAPGKGL

EWIGFINTGGTIVYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARGSYVSSGYAYYFN

V (SEQ ID NO: 443).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 444; SEQ ID NO: 445 and SEQ ID NO: 446 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 442 and / or one or more of the polypeptide sequences from SEQ ID NO: 447; SEQ ID NO: 448 and SEQ ID NO: 449 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 443 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

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In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 444; SEQ ID NO: 445 and SEQ ID NO: 446 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 442 and / or one or more of the polypeptide sequences from SEQ ID NO: 447; SEQ ID NO: 448 and SEQ ID NO: 449 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 443 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 442. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 443.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 444; SEQ ID NO: 445 and SEQ ID NO: 446 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 442.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 447; SEQ ID NO: 448 and SEQ ID NO: 449 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 443.

The invention also contemplates antibody fragments that include one or more antibody fragments described herein. In one embodiment of the invention, antibody fragments possessing IL-6 binding specificity comprise or alternatively consist of one, two, three or more antibody fragments, including all of the following: the strand region light variable of SEQ ID NO: 442, the variable heavy chain region of SEQ ID NO: 443, the complementarity determining regions (SEQ ID NO: 444; SEQ ID NO: 445 and SEQ ID NO: 446) of the variable light chain region of SEQ ID NO: 442 and the regions

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MEXICAN INSTITUTE OF INDUSTRIAL E RCOAD INDUSTRIAL DETERMINANTS OF COMPLEMENTARITY (SEQ ID NO: 447; SEQ ID NO:

448 and SEQ ID NO: 449) of the variable heavy chain region of

<td colspan="2">In a</td><td>modality</td><td>preferred</td><td>of</td><td>the invention, the</td><td>antibody</td>

<td>anti-IL-6</td><td>is</td><td>Ab28 that</td><td>understands</td><td>the</td><td>SEQ ID NO: 442 and</td><td>the SEQ ID</td>

<td>NO: 443,</td><td>than</td><td>owns the</td><td>minus one</td><td>of</td><td>the activities</td><td>biological</td>

established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQSSQSVYNNNWLSWFQQK

PGQPPKLLIYKASTLASGVPSRFKGSGSGTQFTLTISDVQCDDVATYYCAGGYLDSVI (SEQ

ID NO: 458)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSTYSINWVRQAPGKGL

EWIGIIANSGTTFYANWAKGRFTVSKTSTTVDLKITSPTTEDTATYFCARESGMYNEYGKFNI (SEQ ID NO: 459).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 460; SEQ ID NO: 461 and SEQ ID NO: 462 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

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SEQ ID NO: 458 and / or one or more of the polypeptide sequences of SEQ ID NO: 463; SEQ ID NO: 464 and SEQ ID NO: 465 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 459 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 460; SEQ ID NO: 461 and SEQ ID NO: 462 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 458 and / or one or more of the polypeptide sequences from SEQ ID NO: 463; SEQ ID NO: 464 and SEQ ID NO: 465 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 459 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody are also contemplated by the invention.

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY possessing specificity for binding to IL-6. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 458. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 459.

461 and SEQ ID NO: 462 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 458.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 463; SEQ ID NO:

464 and SEQ ID NO: 465 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 459.

The invention also contemplates antibody fragments that include one or more antibody fragments described in

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ΙΝ ^ ΤΓ, ί; ·! ο

LA Η present. In one embodiment of the invention, antibody fragments possessing IL ^ * 6 binding specificity comprise or alternatively consist of one, two, three, or more antibody fragments, including all of the following: the region of SEQ ID variable light chain

NO: 458, the variable heavy chain region of SEQ ID NO:

459, the complementarity determining regions (SEQ ID

NO: 460; SEQ ID NO: 461 and SEQ ID NO: 462) of the variable light chain region of SEQ ID NO: 458 and the complementarity determining regions (SEQ ID NO: 463; SEQ ID NO: 464 and SEQ ID NO: 465 ) of the variable heavy chain region of SEQ ID NO: 459.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab29 comprising SEQ ID NO: 458 and SEQ ID

NO: 459, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCASDMTQTPSSVSAAVGGTVTINCQASENIYSFLAWYQQKPG

QPPKLLIFKASTLASGVSSRFKGSGSGTQFTLTISDLECDDAATYYCQQGATVYDIDNN (SEQ ID NO: 474)

The invention also includes antibodies that possess IL-6 binding specificity and a heavy chain sequence.

191

ΙΜΡΐ

INSTITUTO MEXICANO ce the variable property that comprises the sequence established below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGÍDLSAYAMIWVKUTrPGEGL

EWITIIYPNGITYYANWAKGRFTVSKTSTAMDLKITSPTTEDTATYFCARDAESSKNAYWGYF

NV (SEQ ID NO: 475).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID.

NO: 476; SEQ ID NO: 477 and SEQ ID NO: 478 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 474 and / or one or more of the polypeptide sequences of SEQ ID NO: 479; SEQ ID NO: 480 and SEQ ID NO: 481 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 475 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 476; SEQ ID NO: 477 and SEQ ID NO: 478 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 47 4 and / or one or more of the polypeptide sequences

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INDUSTRIAL

from SEQ ID NO: 479; SEQ ID NO: 480 and SEQ ID NO: 481 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 475 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 474. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 475.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 476; SEQ ID NO: 477 and SEQ ID NO: 478 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 474.

In a further embodiment of the invention, ios

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'Ϊ'; “Ό MEXICAN INDIVIDUAL PROPERTY

Fragments of the antibody possessing IL-6 binding specificity comprise or alternatively consist of one or more polypeptide sequences of SEQ ID NO: 479; SEQ ID NO:

480 and SEQ ID NO: 481 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 475.

NO: 474, the variable heavy chain region of SEQ ID NO:

475, the complementarity determining regions (SEQ ID NO: 476; SEQ ID NO: 477 and SEQ ID NO: 478) of the variable light chain region of SEQ ID NO: 474 and the complementarity determining regions (SEQ ID NO: : 479; SEQ ID NO: 480 and SEQ ID NO: 481) of the variable heavy chain region of SEQ ID NO: 475.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab30 comprising SEQ ID NO: 474 and SEQ ID

NO: 475, which has at least one of the biological activities established herein.

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In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDT RAPTQLLGLLLLWL PGARCAS DMTQT PS SVSAAVGGTVTINCQASENIYS FLAWYQQKPG

QPPKLLIFRASTLASGVSSRFKGSGSGTQFTLTISDLECDDAATYYCQQGATVYDIDNN (SEQ ID NO: 490)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGIDLSAYAMIWVRQAPGEGL

EWITIIYPNGITYYANWAKGRFTVSKTSTAMDLKITSPTTEDTATYFCARDAESSKNAYWGYF

NV (SEQ ID NO: 491).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 492; SEQ ID NO: 493 and SEQ ID NO: 494 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 490 and / or one or more of the polypeptide sequences from SEQ ID NO: 495; SEQ ID NO: 496 and SEQ ID NO: 497 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 491 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 492; SEQ ID NO: 493 and SEQ ID NO: 494 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 490 and / or one or more of the polypeptide sequences from SEQ ID NO: 495; SEQ ID NO: 496 and SEQ ID NO: 497 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 491 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 490. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 491.

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In a further embodiment of the invention, Tos fragments of the antibody possessing binding specificity=IL-6 comprises or alternatively consists of one or more polypeptide sequences of SEQ ID NO: 492; SEQ ID NO:

493 and SEQ ID NO: 494 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 490.

496 and SEQ ID NO: 497 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 491.

NO: 490, the variable heavy chain region of SEQ ID NO: 491, the complementarity determining regions (SEQ ID

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

NO: 492; SEQ ID NO: 493 and SEQ ID NO: 494) of the variable light chain region of SEQ ID NO: 490 and the complementarity determining regions (SEQ ID NO: 495; SEQ ID NO:

496 and SEQ ID NO: 497) of the variable heavy chain region of SEQ ID NO: 491.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab31 comprising SEQ ID NO: 490 and SEQ ID NO: 491, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAIEMTQTPSPVSAAVGGTVTINCQASESVFNNMLSWYQQKP

GHSPKLLIYDASDLASGVPSRFKGSGSGTQFTLTISGVECDDAATYYCAGYKSDSNDGDNV (SEQ ID NO: 506)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLNRNSITWVRQAPGEGL

EWIGIITGSGRTYYANWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARGHPGLGSGNI (SEQ ID NO: 507).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 508; SEQ ID NO: 509 and SEQ ID NO: 510 which correspond to the

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL Complementarity determining regions (CDRs or hypervariables) of the variable light chain sequence of SEQ ID NO: 506 and / or one or more of the polypeptide sequences of SEQ ID NO: 511; SEQ ID NO: 512 and SEQ ID NO: 513 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 507 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 508; SEQ ID NO: 509 and SEQ ID NO: 510 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 506 and / or one or more of the polypeptide sequences from SEQ ID NO: 511; SEQ ID NO: 512 and SEQ ID NO: 513 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 507 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable light and heavy chain sequences

199

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MEXICAN INSTITUTE OF PROPERTY previously established. industrial

The invention also contemplates fragilli liLUy dei -'- antiene ^ po · possessing IL-6 binding specificity. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 506. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 507.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 508; SEQ ID NO: 509 and SEQ ID NO: 510 which correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 506.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 511; SEQ ID NO:

512 and SEQ ID NO: 513 that correspond to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 507.

200

NO: 506, the variable heavy chain region of SEQ ID NO:

507, the complementarity determining regions (SEQ ID

NO: 508; SEQ ID NO: 509 and SEQ ID NO: 510) of the variable light chain region of SEQ ID NO: 506 and the complementarity determining regions (SEQ ID NO: 511; SEQ ID NO: 512 and SEQ ID NO: 513 ) of the variable heavy chain region of SEQ ID NO: 507.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab32 comprising SEQ ID NO: 506 and SEQ ID

NO: 507, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAQVLTQTASSVSAAVGGTVTINCQSSQSVYNNYLSWYQQKP

GQPPKLLIYTASSLASGVPSRFKGSGSGTQFTLTISEVQCDDAATYYCQGYYSGPIIT (SEQ

ID NO: 522)

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IMPI

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLNNYYIQWVRQAPGEGL

EWIGIIYAGGSAYYATWANGRFTIAKTSSTTVDLKMTSLTTEDTATYFCARGTFDGYEL (SEQ ID NO: 523).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 524; SEQ ID NO: 525 and SEQ ID NO: 526 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 522 and / or one or more of the polypeptide sequences from SEQ ID NO: 527; SEQ ID NO: 528 and SEQ ID NO: 529 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 523 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 524; SEQ ID NO: 525 and SEQ ID NO: 526 corresponding to the complementarity determining regions (CDRs or regions

202

IMPI

hypervariables) of the light chain sequence'variable

SEQ ID NO: 522 and / or one or more of the poly ^ 'é ^ T'idós sequences of SEQ ID NO: 527; SEQ ID NO: 528 and SEQ ID NO: 529 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 523 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 522. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 523.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 524; SEQ ID NO: 525 and SEQ ID NO: 526 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

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SEQ ID NO: 522.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 527; SEQ ID NO: 528 and SEQ ID NO: 529 corresponding to Complementarity-determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 523.

NO: 522, the variable heavy chain region of SEQ ID NO: 523, the complementarity determining regions (SEQ ID

NO: 524; SEQ ID NO: 525 and SEQ ID NO: 526) of the variable light chain region of SEQ ID NO: 522 and the complementarity determining regions (SEQ ID NO: 527; SEQ ID NO: 528 and SEQ ID NO: 529 ) of the variable heavy chain region of SEQ ID NO: 523.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab33 comprising SEQ ID NO: 522 and SEQ ID

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MEXICAN INSTITUTE OF INDUSTAAL PÍOXIEDAD

NO: 523, which has at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSPVSVPVGDTVTISCQSSESVYSNNLLSWYQQK

PGQPPKLLIYRASNLASGVPSRFKGSGSGTQFTLTISGAQCDDAATYYCQGYYSGVINS (SEQ ID NO: 538)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMSWVRQAPGEGL

EYIGFINPGGSAYYASWASGRLTISKTSTTVDLKITSPTTEDTATYFCARILIVSYGAFTI (SEQ ID NO: 539).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 540; SEQ ID NO: 541 and SEQ ID NO: 542 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 538 and / or one or more of the polypeptide sequences from SEQ ID NO: 543; SEQ ID NO: 544 and SEQ ID NO: 545 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 539 or combinations thereof

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polypeptide sequences. In another embodiment of the invention .. the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID NO: 540; SEQ ID NO: 541 and SEQ ID NO: 542 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 538 and / or one or more of the polypeptide sequences from SEQ ID NO: 543; SEQ ID NO: 544 and SEQ ID NO: 545 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 539 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 538. In another embodiment of the invention, the antibody fragments of the invention

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IMPI Mexican iwstítuw O £ LA fSíWDAÜ LXSU2TS1AL

comprise, or alternatively consist of, a polypeptide sequence of SEQ ID NO: 539.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 540; SEQ ID NO:

541 and SEQ ID NO: 542 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 538.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 543; SEQ ID NO:

544 and SEQ ID NO: 545 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 539.

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OF THE PROPERTY, _ INDUSTRIAL _ _

NO: 538, the variable heavy chain region of SEQ ID NO:

539, the complementarity determining regions (SEQ ID

NO: 540; SEQ ID NO: 541 and SEQ ID NO: 542) of the variable light chain region of SEQ ID NO: 538 and the complementarity determining regions (SEQ ID NO: 543; SEQ ID NO:

544 and SEQ ID NO: 545) of the variable heavy chain region of SEQ ID NO: 539.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab34 comprising SEQ ID NO: 538 and SEQ ID NO: 539, which possesses at least one of the biological activities established herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQATESIGNELSWYQQKPG

QAPKLLIYSASTLASGVPSRFKGSGSGTQFTLTITGVECDDAATYYCQQGYSSANIDNA (SEQ ID NO: 554)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSKYYMSWVRQAPEKGL KYIGYIDSTTVNTYYATWARGRjFTISKTSTTVDLKITSPTSEDTATYFCARGSTYFTDGGHRL DL (SEQ ID NO: 55

The invention further contemplates antibodies that

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they comprise one or more polypeptide sequences of SEQ ID NO: 556; SEQ ID NO: 557 and SEQ ID NO: 558 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 554 and / or one or more of the polypeptide sequences from SEQ ID NO: 559; SEQ ID NO: 560 and SEQ ID NO: 561 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 555 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

NO: 556; SEQ ID NO: 557 and SEQ ID NO: 558 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 554 and / or one or more of the polypeptide sequences from SEQ ID NO: 559; SEQ ID NO: 560 and SEQ ID NO: 561 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 555 or combinations of these polypeptide sequences. In another embodiment of the invention,

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY !.

the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 554. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 555.

In a further embodiment of the invention, fragments of the antibody possessing IL-6 binding specificity comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 556; SEQ ID NO: 557 and SEQ ID NO: 558 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 554.

560 and SEQ ID NO: 561 corresponding to the complementarity determining regions (CDR or regions

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INSTITUTO MEXICANO DE LA PROPERTY INDUSTRIAL hipervariables) of the variable heavy chain sequence of the

SEQ ID NO: 555.

<td>NOT:</td><td> 554,</td><td>the region</td><td>Variable Heavy Chain SEQ ID NO:</td>

<td> 555,</td><td>the</td><td>regions</td><td>determinants of complementarity (SEQ ID</td>

<td>NOT:</td><td> 556;</td><td colspan="2">SEQ ID NO: 557 and SEQ ID NO: 558) of the region of</td>

variable light chain of SEQ ID NO: 554 and complementarity determining regions (SEQ ID NO: 559; SEQ ID NO:

560 and SEQ ID NO: 561) of the variable heavy chain region of SEQ ID NO: 555.

In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab35 comprising SEQ ID NO: 554 and SEQ ID NO: 555, which possesses at least one of the biological activities set forth herein.

In another embodiment, the invention includes antibodies possessing IL-6 binding specificity and a variable light chain sequence comprising the sequence set forth below:

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQATESIGNELSWYQQKPG

211

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INSTITUTO MEXíCANC DE LA HROPICDA!

QAPKLLIYSASTLASGVPSRFKGSGSGTQFTLTITGVECDDAATYYCQW ^ ANI (SEQ ID NO: 570)

The invention also includes antibodies possessing IL-6 binding specificity and a variable heavy chain sequence comprising the sequence set forth below:

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSTYNMGWVRQAPGKGL

EWIGSITIDGRTYYASWAKGRFTVSKSSTTVDLKMTSLTTGDTATYFCARILIVSYGAFTI (SEQ ID NO: 571).

The invention further contemplates antibodies comprising one or more polypeptide sequences of SEQ ID NO: 572; SEQ ID NO: 573 and SEQ ID NO: 574 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 570 and / or one or more of the polypeptide sequences from SEQ ID NO: 575; SEQ ID NO: 576 and SEQ ID NO: 577 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 571 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the

CDR and the variable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric antibodies, comprising one or more polypeptide sequences of SEQ ID

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NO: 572; SEQ ID NO: 573 and SEQ ID NO: 574 corresponding to the complementarity determining regions (CDRs or 'hypervariable regions') of the variable light chain sequence of SEQ ID NO: 570 and / or one or more of the sequences of polypeptides of SEQ ID NO: 575; SEQ ID NO: 576 and SEQ ID NO: 577 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 571 or combinations of these polypeptide sequences. In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above.

Fragments of the antibody possessing IL-6 binding specificity are also contemplated by the invention. In one embodiment of the invention, the antibody fragments of the invention comprise, or alternatively consist of, the polypeptide sequence of SEQ ID NO: 570. In another embodiment of the invention, the antibody fragments of the invention comprise , or alternatively consist of a polypeptide sequence of SEQ ID NO: 571.

573 and SEQ ID NO: 574 corresponding to the regions

213 hypervariable determinants) of chain sequence complementarity

INSTITUTO MEXICANO DE LA PR '/ PIEDAD INDUSTRIAL (CDR or light regions variáBTe ™ 3e ^ a ”

SEQ ID NO: 570.

In a further embodiment of the invention, fragments of the antibody possessing binding specificity for

IL-6 comprise or, alternatively, consist of one or more polypeptide sequences of SEQ ID NO: 575; SEQ ID NO:

576 and SEQ ID NO: 577 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 571.

<td>NOT:</td><td> 570,</td><td>the region</td><td colspan="2">heavy chain</td><td>SEQ ID variable</td><td>NOT:</td>

<td> 571,</td><td>the</td><td>regions</td><td>determinants</td><td>of</td><td>complementarity (SEQ</td><td>ID</td>

<td>NOT:</td><td> 572;</td><td colspan="2">SEQ ID NO: 573 and SEQ</td><td>ID</td><td>NO: 57 4) from the region</td><td>of</td>

variable light chain of SEQ ID NO: 570 and the complementarity determining regions (SEQ ID NO: 57 5; SEQ ID NO: 576 and SEQ ID NO: 577) of the variable heavy chain region of SEQ ID NO: 571 .

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In a preferred embodiment of the invention, the anti-IL-6 antibody is Ab36 comprising SEQ ID NO: 570 and SEQ ID

NO: 571, which has at least one of the biological activities established herein.

The anti-IL-6 antibody sequences of the present invention are shown in Table 1. Examples of sequence variants and other alternative forms of the heavy and light chains from Abl to Ab7 are shown. The antibodies of the present invention comprise additional sequence variants that include conservative substitutions, substitutions of one or more CDR sequences and / or sequences of

FR etc.

Examples of Abl modalities include an antibody comprising a light chain and / or heavy chain variant. Examples of Abl light chain variants include the sequence of any of the Abl light chains.

Abl shown (i.e. any of SEQ ID NO: 2, 20, 647, 651, 660, 666, 699, 702, 706 or 709) where the entire CDR1 sequence is replaced or where one is substituted or more residues in the CDRl sequence with the residue in the corresponding position of any of the other established light chain CDRl sequences (i.e., any of SEQ ID NO: 23, 39, 55, 71, 87, 103, 124 , 140, 156, 172, 188, 204,

220, 236, 252, 268, 284, 300, 316, 332, 348, 364, 380, 396,

412, 428, 444, 460, 476, 492, 508, 524, 540, 556 or 572) and / or

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CS THE PROPERTY L The - ..,,

INDUSTRIAL ^ «« to Ui where the entire CDR2 sequence is replaced or where one or more residues in the CDR2 sequence is replaced with “the” residue at the corresponding position of any of the other established light chain CDR2 sequences (i.e. any of SEQ ID NO: 24, 40, 56, 72, 88, 104,

125, 141, 157, 173, 189, 205, 221, 237, 253, 269, 285, 301,

317, 333, 349, 365, 381, 397, 413, 429, 445, 461, 477, 493,

509, 525, 541, 557 or 573) and / or where the entire CDR3 sequence is replaced or where one or more residues in the CDR3 sequence is replaced with the residue at the corresponding position of any of the other sequences of Established light chain CDR3s (i.e. any of the SEQ

ID NO: 25, 41, 57, 73, 89, 105, 126, 142, 158, 174, 190, 206,

222, 238, 254, 270, 286, 302, 318, 334, 350, 366, 382, 398,

414, 430, 446, 462, 478, 494, 510, 526, 542, 558, or 574).

Examples of Abl heavy chain variants include the sequence of any of the Abl heavy chains shown (i.e., any of SEQ ID NO: 3,

18, 19, 652, 656, 657, 658, 661, 664, 665, 704 or 708) where the entire CDR1 sequence is replaced or where one or more residues in the CDR1 sequence is replaced with the residue in the corresponding position of any of the other established heavy chain CDRl sequences (i.e. any of SEQ ID NO: 26, 42, 58, 74, 90, 106,

127, 143, 159, 175, 191, 207, 223, 239, 255, 271, 287, 303,

216

511, 527, 543, 559, or 575) and / or where the entire CDR2 sequence is replaced or where one or more residues in the CDR2 sequence is replaced with the residue at the corresponding position on a heavy chain CDR2 of Abl, as set forth in Table 1 (i.e. any of SEQ ID NO:

or 120) or any of the other established heavy chain CDR2 sequences (i.e., any of SEQ ID NO: 27,

43, 59, 75, 91, 107, 121, 128, 144, 160, 176, 192, 208, 224,

240, 256, 272, 288, 304, 320, 336, 352, 368, 384, 400, 416, 432, 448, 464, 480, 496, 512, 528, 544, 560 or 576) and / or where it is replaced the entire CDR3 sequence or where one or more residues in the CDR3 sequence is replaced with the residue at the corresponding position of any of the other established heavy chain CDR3 sequences (is

513, 529, 545, 561 or 577).

In another embodiment, the invention contemplates other antibodies such as, for example, chimeric or humanized antibodies comprising one or more polypeptide sequences of SEQ ID NO: 4; SEQ ID NO: 5 and SEQ ID NO: 6 corresponding to the complementarity determining regions (CDRs or hypervariable regions) of the chain sequence

217

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY light variable SEQ ID NO: 2 and / or one or more of the polypeptide sequences of SEQ ID NO: 7 (CDR1); SEQ ID

NO: 8 (CDR2); SEQ ID NO: 120 (CDR2); and SEQ ID NO: 9 (CDR3) corresponding to the complementarity determining regions (CDR or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO: 3 or SEQ ID NO: 19 or combinations of these polypeptide sequences . In another embodiment of the invention, the antibodies of the invention include combinations of the CDRs and the variable heavy and light chain sequences set forth above, including those set forth in Figures: 2 and 34-37, as well as those identified in Table 1 .

In another embodiment, the anti-IL-6 antibody of the invention is one comprising at least one of the following: a CDR1 light chain encoded by the sequence in SEQ ID NO: 12 or SEQ ID NO: 694, a light chain of CDR2 encoded by the sequence in SEQ ID NO: 13, a CDR3 light chain encoded by the sequence in SEQ ID NO: 14 or SEQ ID NO: 695, a heavy chain of CDR1 encoded by the sequence in SEQ ID

NO: 15, a CDR2 heavy chain encoded by SEQ ID NO: 16 or SEQ ID NO: 696 and a CDR3 heavy chain encoded by SEQ ID NO: 17 or SEQ ID NO: 697. Additionally, the invention encompasses such nucleic acid sequences and variants thereof.

In another embodiment, the invention is directed to sequences of

218

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY amino acids that correspond to the CDRs of said anti-IL-6 antibody that are selected from SEQ ID NO: 4 (CDR1), SEQ ID

NO: 5 (CDR2), SEQ ID NO: 6 (CDR3), SEQ ID NO: 7, SEQ ID NO: 120 and SEQ ID NO: 9.

In another embodiment, the anti-IL-6 antibody of the invention comprises a light chain nucleic acid sequence of SEQ ID NO: 10, 662, 698, 701, 705, 720, 721, 722 or 723 and / or a heavy chain nucleic acid sequence of

SEQ ID NO: 11, 663, 700, 703, 707, 724 or 725. Additionally, the invention is directed to the corresponding polypeptides encoded by any of the above nucleic acid sequences and combinations thereof.

In a specific embodiment of the invention, anti-IL-6 antibodies or a portion thereof will be encoded by a nucleic acid sequence selected from those comprised in SEQ ID NO: 10, 12, 13, 14, 662, 694, 695,

698, 701, 705, 720, 721, 722, 723, 11, 15, 16, 17, 663, 696,

697, 700, 703, 707, 724 and 725. For example, CDR1 on the light chain may be encoded by SEQ ID NO: 12 or

694, CDR2 on the light chain can be encoded by SEQ ID NO: 13, CDR3 on the light chain can be encoded by SEQ ID NO: 14 or 695, CDR1 on the heavy chain can be encoded by SEQ ID NO: 15, CDR2 in the heavy chain may be encoded by SEQ ID NO: 16 or

696, CDR3 in the heavy chain may be encoded by

219

IMPI

INSTITUTE .'- 'f' / ICANO £ 1 LA

SEQ ID NO: 17 or 697. As discussed at confirmation! Antibodies containing these CDRs can be created using appropriate human flanking regions based on the humanization methods described herein.

In another specific embodiment of the invention, the variable light chain will be encoded by SEQ ID NO: 10, 662,

698, 701, 705, 720, 721, 722 or 723 and the variable heavy chain of anti-IL-6 antibodies will be encoded by SEQ ID

NO: 11, 663, 700, 703, 707, 724 or 725.

In a more specific embodiment, the variable heavy and light chains of the anti-IL-6 antibody will be encoded by SEQ ID NO: 10 and 11 or SEQ ID NO: 698 and SEQ ID NO: 700 or SEQ ID NO: 701 and SEQ ID NO: 703 or SEQ ID NO: 705 and SEQ

ID NO: 707.

In another specific embodiment, the invention encompasses nucleic acid constructs containing any of the preceding nucleic acid sequences and combinations thereof, as well as recombinant cells containing these nucleic acid constructs and sequences, where these nucleic acid constructs or sequences. they can be extrachromosomal or they can be integrated into the genome of the host cell.

In another specific embodiment, the invention encompasses polypeptides containing any of the CDRs or combinations thereof described in SEQ ID NO: 4, SEQ

220

IMPI

ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ TT5NO: 120, SEQ ID NO: 9 or polypeptides comprising any of the variable light chain polypeptides comprised in SEQ ID NO: 2, 20, 647, 651, 660, 666, 699, 702, 706 or 709 and / or the variable heavy chain polypeptides included in

SEQ ID NO: 3, 18, 19, 652, 656, 657, 658, 661, 664, 665, 704 or

708. Optionally, these polypeptides can be linked, directly or indirectly, to other immunogiobulin polypeptides or effector moieties, such as therapeutic or detectable entities.

In another embodiment, the anti-IL-6 antibody is one comprising at least one of the following: a variable light chain encoded by the sequence in SEQ ID NO: 10 or SEQ ID

NO: 698 or SEQ ID NO: 701 or SEQ ID NO: 705 and a variable string encoded by the sequence in SEQ ID NO: 11 or SEQ ID NO: 700 or

SEQ ID NO: 703 or SEQ ID NO: 707.

In another embodiment, the anti-IL-6 antibody is a variant of the preceding sequences that includes one or more substitutions on the flanking and / or CDR sequences and that has one or more of the properties of Abl in vitro and / or subsequently of in vivo administration.

These in vitro and in vivo properties are described in more detail in the examples below and include: competing with Abl for binding to IL-6 and / or peptides thereof, having a binding affinity (Kd) for IL-6, or less than about 50

221

IMPI

picomolar and / or a dissociation rate (Koff) IL-6 less than or equal to 10 ~4 S_1, have an in vivo half-life of at least about 22 days in a healthy human being, have the ability to prevent or treat hypoalbuminemia, the ability to prevent or treat elevated CRP, the ability to prevent or treat abnormal coagulation and / or the ability to decrease the risk of thrombosis in an individual with a disease or condition associated with the increased risk of thrombosis.

Additional non-limiting examples of anti-IL-6 activity are set forth herein, for example, under the title

Anti-IL-6 activity.

In another embodiment, the anti-IL-6 antibody includes one or more of the Abl light and / or heavy chain CDR sequences (see Table 1) or a variant or variants thereof having / n one or more of Abl properties in vitro and / or after in vivo administration (examples of such properties are discussed in the previous paragraph). One skilled in the art would understand how to combine these CDR sequences to form an antigen-binding surface, for example, by binding to one or more scaffolds that may comprise human or other mammalian flanking sequences or their functional orthologs that come from a SMIP, camelid antibody, nanoantibody, IgNAR, or other immunoglobulin or other designed antibody. For example, the modalities can specifically bind human IL-6 and include one, two, three,

222

IMP four, five, six or more of the following CDR sequences or variants thereof:

a polypeptide having at least a 72.7% identity (ie 8 out of 11 amino acids) with the CDR1 light chain of SEQ ID NO: 4;

a polypeptide having at least an 81.8% identity (ie, 9 out of 11 amino acids) with the CDR1 light chain of SEQ ID NO: 4;

a polypeptide having at least 90.9% identity (ie, 10 out of 11 amino acids) with the CDR1 light chain of SEQ ID NO: 4;

a polypeptide that has at least 100% identity (ie, 11 out of 11 amino acids) with the CDR1 light chain of SEQ ID NO: 4;

a polypeptide having at least an 85.7% identity (ie 6 out of 7 amino acids) with the CDR2 light chain of SEQ ID NO: 5;

a polypeptide having at least 100% identity (ie, 7 out of 7 amino acids) with the CDR2 light chain of SEQ ID NO: 5;

a polypeptide having at least 50% identity (ie 6 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polypeptide having at least 58.3% identity (i.e., 7 out of 12 amino acids) with the CDR3 light chain of

223

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

SEQ ID NO: 6;

a polypeptide having at least a 66.6% identity (ie 8 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polypeptide that has at least 75% identity (ie 9 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polypeptide possessing at least 83.3% identity (ie 10 out of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polypeptide having at least a 91.6% identity (ie, 11 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polypeptide that has at least 100% identity (ie, 12 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polypeptide having at least 80% identity (ie, 4 out of 5 amino acids) with the CDR1 heavy chain of SEQ ID NO: 7;

a polypeptide having at least 100% identity (ie, 5 out of 5 amino acids) with the CDR1 heavy chain of SEQ ID NO: 7;

a polypeptide having at least 50% identity (ie 8 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

224

IMPI

a polypeptide possessing at least 56.2% identity (ie, 9 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polypeptide possessing al (i.e. 10 out of 16 amino acids) of SEQ ID NO: 120;

a polypeptide possessing al (i.e., 11 out of 16 amino acids) of SEQ ID NO: 120;

a polypeptide possessing al (i.e., 12 of 16 amino acids) of SEQ ID NO: 120;

a polypeptide possessing al (ie 13 of 16 amino acids) of SEQ ID NO: 120;

a polypeptide possessing al (i.e., 14 out of 16 amino acids) of SEQ ID NO: 120;

a polypeptide possessing al (i.e., 15 of 16 amino acids) of SEQ ID NO: 120;

a polypeptide possessing al (i.e. 16 out of 16 amino acids) of SEQ ID NO: 120;

a polypeptide having at least 62.5% identity with CDR2 heavy chain minus 68.7% identity with CDR2 heavy chain minus 75% identity with CDR2 heavy chain minus 81 identity , 2% with CDR2 heavy chain minus 87.5% identity with CDR2 heavy chain minus 93.7% identity with CDR2 heavy chain minus 100% identity with CDR2 heavy chain minus identity 33.3%

225

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY (ie, 4 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9; * a polypeptide having at least 41.6% identity (ie, 5 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide having at least 50% identity (ie 6 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide having at least 58.3% identity (ie 7 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide having at least a 66.6% identity (ie 8 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide having at least 75% identity (ie, 9 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide possessing at least 83.3% identity (ie, 10 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide having at least a 91.6% identity (ie, 11 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide that has at least 100% identity (i.e., 12 of 12 amino acids) with the CDR3 heavy chain

226

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

from SEQ ID NO: 9;

a polypeptide that is at least 90.9% similar (ie 10 out of 11 amino acids) to the CDRl light chain of SEQ ID NO: 4;

a polypeptide that is 100% similar (ie 11 out of 11 amino acids) to the CDRl light chain of SEQ ID

NO: 4;

a polypeptide that is at least 85.7% similar (ie 6 out of 7 amino acids) to the CDR2 light chain of SEQ ID NO: 5;

a polypeptide that is at least 100% similar (ie 7 out of 7 amino acids) to the CDR2 light chain of SEQ ID NO: 5;

a polypeptide that is at least 66.6% similar (ie 8 out of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

a polypeptide that is at least 75% similar (i.e. 9 out of 12 amino acids) to the CDR3 light chain of the

SEQ ID NO: 6;

a polypeptide that is at least 83.3% similar (ie 10 out of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

a polypeptide that is at least 91.6% similar (ie 11 out of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

227

¥ ti

JK.

INSTITUTO MEXICANO DE LA VF.

NO 6;

a polypeptide that is at least 80% similar (i.e. 4 out of 5 amino acids) to the CDR1 heavy chain of the

SEQ ID NO: 7;

a polypeptide that is at least 100% similar (i.e. 5 out of 5 amino acids) to the CDR1 heavy chain of the

SEQ ID NO: 7;

a polypeptide that is at least 56.2% similar (ie 9 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polypeptide that is at least 62.5% similar (i.e. 10 out of 16 amino acids) to the CDR2 heavy chain of the

SEQ ID NO: 120;

a polypeptide that has at least 68.7% similarity (i.e. 11 out of 16 amino acids) to the CDR2 heavy chain of the

SEQ ID NO: 120;

a polypeptide that bears at least 75% similarity (i.e. 12 out of 16 amino acids) to the CDR2 heavy chain of the

SEQ ID NO: 120;

a polypeptide that has at least 81.2% similarity (i.e., 13 of 16 amino acids) to the CDR2 heavy chain of the

SEQ ID NO: 120;

a polypeptide that has at least 87.5% similarity (is

228

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY (14 of 16 amino acids) with the CDR2 heavy chain of the

SEQ ID NO: 120;

a polypeptide that is at least 93.7% similar (i.e. 15 out of 16 amino acids) to the CDR2 heavy chain of the

SEQ ID NO: 120;

a polypeptide that has 100% similarity (i.e. 16 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID

NO: 120;

a polypeptide that has at least 50% similarity (i.e. 6 out of 12 amino acids) to the CDR3 heavy chain of the

SEQ ID NO: 9;

a polypeptide that is at least 58.3% similar (ie 7 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide that is at least 66.6% similar (ie 8 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide that is at least 75% similar (i.e. 9 out of 12 amino acids) to the CDR3 heavy chain of the

SEQ ID NO: 9;

a polypeptide that is at least 83.3% similar (ie 10 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

a polypeptide that has at least 91.6% similarity (i.e. 11 out of 12 amino acids) to the CDR3 heavy chain

229 from SEQ ID NO: 9;

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

a polypeptide that has 100% similarity (ie 12 of 12 amino acids) to the CDR3 heavy chain of SEQ ID

NO: 9.

Other examples of modalities include one or more polynucleotides that encode any of the above, for example, a polynucieotide that encodes a polypeptide that specifically binds human IL-6 and includes one, two, three, four, five, six or more of the following CDRs or variants thereof:

a polynucieotide encoding a polypeptide having at least 72.7% identity (ie 8 out of 11 amino acids) with the CDR1 light chain of SEQ ID NO: 4;

a polynucieotide encoding a polypeptide having at least 81.8% identity (ie 9 out of 11 amino acids) with the CDR1 light chain of SEQ ID NO: 4;

a polynucieotide encoding a polypeptide having at least 90.9% identity (ie, 10 out of 11 amino acids) with the CDRl light chain of SEQ ID NO: 4;

a polynucieotide encoding a polypeptide with 100% identity (ie 11 out of 11 amino acids) with the CDRl light chain of SEQ ID NO: 4;

a polynucieotide encoding a polypeptide with at least 85.7% identity (ie 6 out of 7 amino acids) with the CDR2 light chain of SEQ ID NO: 5;

230

IMPI

INSTITUTO MEXICANO CE LA PROPIEDAD INDUSTRIAL a polynucleotide that encodes a polypeptide that has 100% identity (ie 7 out of 7 amino acids) with the CDR2 light chain of SEQ ID NO: 5;

a polynucleotide encoding a polypeptide having at least 50% identity (ie 6 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide having at least 58.3% identity (ie 7 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide having at least 66.6% identity (ie 8 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide having at least 75% identity (ie 9 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide having at least 83.3% identity (ie, 10 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide having at least 91.6% identity (ie, 11 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide that encodes a polypeptide that has

100% identity (ie 12 of 12 amino acids) with the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide that encodes a polypeptide that has

231

ΙΜΡΐ

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

at least 80% identity (ie 4 out of 5 amino acids) with the CDR1 heavy chain of SEQ ID NO: 7; "" A polynucleotide encoding a polypeptide having 100% identity (ie 5 out of 5 amino acids) with the CDR1 heavy chain of SEQ ID NO: 7;

a polynucleotide encoding a polypeptide having at least 50% identity (ie 8 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide having at least 56.2% identity (ie, 9 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide having at least 62.5% identity (ie, 10 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide having at least 68.7% identity (ie, 11 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide having at least 75% identity (ie 12 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide having at least 81.2% identity (ie 13 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide that encodes a polypeptide that has at least 87.5% identity (i.e., 14 out of 16 amino acids)

232

IMPI

MEXICAN INSTITUTE OF PROPERTY with the CDR2 heavy chain of SEQ ID NO: i¿0AND/sn <1AL a polynucleotide encoding a Jjue ETéñé · polypeptide at least 93.7% identity (ie 15 of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide that encodes a polypeptide that has

100% identity (ie 16 out of 16 amino acids) with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide having at least 33.3% identity (ie, 4 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide having at least 41.6% identity (ie, 5 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide having at least 50% identity (ie 6 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide having at least 58.3% identity (ie, 7 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide having at least 66.6% identity (ie 8 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide having at least 75% identity (ie 9 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

233

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

a polynucleotide encoding a polypeptide having at least 83.3% identity (ie, 10 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide having at least 91.6% identity (ie, 11 of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide that encodes a polypeptide that has

100% identity (ie 12 out of 12 amino acids) with the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide that encodes a polypeptide that is at least 90.9% similar (that is, 10 out of 11 amino acids)

<td colspan="6">with the CDR1 light chain of SEQ ID NO: 4;</td><td rowspan="2">than</td><td rowspan="2">has</td>

<td>a</td><td>polynucleotide</td><td>than</td><td>encode</td><td>a</td><td>polypeptide</td>

<td>100% of</td><td>similarity (is</td><td colspan="2">say 11 of</td><td> 11</td><td colspan="3">amino acids) with the</td>

<td colspan="2">CDR1 light chain</td><td>the</td><td>SEQ ID NO:</td><td> 4;</td><td></td><td></td><td></td>

<td>a</td><td>polynucleotide</td><td>than</td><td>encode</td><td>a</td><td>polypeptide</td><td>than</td><td>has</td>

<td>at least</td><td colspan="2">85.7% similarity</td><td>(that is to say,</td><td> , 6</td><td colspan="3">7 amino acids) with</td>

<td>chain</td><td>CDR2 lightweight</td><td>of</td><td>the SEQ ID</td><td>NOT:</td><td> 5;</td><td></td><td></td>

<td>a</td><td>polynucleotide</td><td>than</td><td>encode</td><td>a</td><td>polypeptide</td><td>than</td><td>has</td>

100% similarity (ie 7 out of 7 amino acids) to the CDR2 light chain of SEQ ID NO: 5;

a polynucleotide encoding a polypeptide that is at least 66.6% similar (ie 8 out of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide that encodes a polypeptide that has

234

at least 75% similarity (ie 9 out of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide that is at least 83.3% similar (ie 10 out of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide that is at least 91.6% similar (ie, 11 of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide that has 100% similarity (ie, 12 of 12 amino acids) to the CDR3 light chain of SEQ ID NO: 6;

a polynucleotide encoding a polypeptide that is at least 80% similar (ie 4 out of 5 amino acids) to the CDR1 heavy chain of SEQ ID NO: 7;

a polynucleotide that encodes a polypeptide that has

100% similarity (ie 5 out of 5 amino acids) to the CDR1 heavy chain of SEQ ID NO: 7;

a polynucleotide encoding a polypeptide that is at least 56.2% similar (ie 9 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide that is at least 62.5% similar (ie 10 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide that encodes a polypeptide that is at least 68.7% similar (that is, 11 out of 16 amino acids)

235

IMPIOUS

INDUSTRIAL with the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide that is at least 75% similar (ie 12 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide that is at least 81.2% similar (ie 13 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide that is at least 87.5% similar (ie 14 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide that is at least 93.7% similar (ie 15 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide that encodes a polypeptide that has

100% similarity (ie 16 out of 16 amino acids) to the CDR2 heavy chain of SEQ ID NO: 120;

a polynucleotide encoding a polypeptide that is at least 50% similar (ie 6 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide that is at least 58.3% similar (ie 7 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide that is at least 66.6% similar (ie 8 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

236

IMPI

INSTITUTO MEXICANO DELA PROPIEDAD a polynucleotide that encodes a polypeptideNicioIA1'what ^'cit has at least 75% similarity (ie, 9 of 'T2 amino amino acids TdS) COTÍ CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide that is at least 83.3% similar (ie 10 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide encoding a polypeptide that is at least 91.6% similar (ie 11 out of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9;

a polynucleotide that encodes a polypeptide that has

100% similarity (i.e. 12 of 12 amino acids) to the CDR3 heavy chain of SEQ ID NO: 9.

Table 1. Example sequences of anti-IL-6 antibodies.

Light chains

Abl *

Antibody

<td>Caden. earlier PRT.</td><td>ace of aerpo Nuc.</td><td>CDl PRT.</td><td>31 Nuc.</td><td>CDl PRT.</td><td>32 Nuc.</td><td>CDl PRT.</td><td>33 Nuc.</td>

237

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<td rowspan="3">Chains light human used in humanization from Abl</td><td> 648</td><td></td><td> 710</td><td></td><td> 713</td><td></td><td></td><td></td>

<td rowspan="9">Chains heavy Abl</td><td> 3</td><td> 11</td><td> 7</td><td> 15</td><td> 8</td><td> 16</td><td> 9</td><td> 17</td>

238 τ>

<td rowspan="4"></td><td> 665</td><td></td><td> 7</td><td> 15</td><td> 120</td><td>STITUTO ME D £ LA PLC 6 <</td><td>JCAN'O ΠΕΡΑ OV ”? S'fRÍAL 9</td><td> 17</td>

<td rowspan="3">Chains heavy human used in the humanization from Abl</td><td> 653</td><td></td><td> 716</td><td></td><td> 717</td><td></td><td></td><td></td>

<td rowspan="2">Chains light Ab2</td><td> 21</td><td> 29</td><td> 23</td><td> 31</td><td> 24</td><td> 32</td><td> 25</td><td> 33</td>

<td rowspan="2">Chains heavy Ab2</td><td> 22</td><td> 30</td><td> 26</td><td> 34</td><td> 27</td><td> 35</td><td> 28</td><td> 36</td>

<td rowspan="2">Chains light Ab3</td><td> 37</td><td> 45</td><td> 39</td><td> 47</td><td> 40</td><td> 48</td><td> 41</td><td> 49</td>

<td rowspan="2">Chains heavy Ab3</td><td> 38</td><td> 46</td><td> 42</td><td> 50</td><td> 43</td><td> 51</td><td> 44</td><td> 52</td>

<td rowspan="2">Chains light Ab4</td><td> 53</td><td> 61</td><td> 55</td><td> 63</td><td> 56</td><td> 64</td><td> 57</td><td> 65</td>

<td>Chains</td><td> 54</td><td> 62</td><td> 58</td><td> 66</td><td> 59</td><td> 67</td><td> 60</td><td> 68</td>

239

IMPI

<td>heavy Ab4</td><td> 676</td><td> 678</td><td> 58</td><td> 66</td><td> 59</td><td>OF THE PP.f IND 67</td><td>PIETY Π ISTRIAL 60</td><td> 68</td>

<td rowspan="2">Chains light of Ab5</td><td> 69</td><td> 77</td><td> 71</td><td> 79</td><td> 72</td><td> 80</td><td> 73</td><td> 81</td>

<td rowspan="2">Chains heavy Ab5</td><td> 70</td><td> 78</td><td> 74</td><td> 82</td><td> 75</td><td> 83</td><td> 76</td><td> 84</td>

<td rowspan="2">Chains light Ab6</td><td> 85</td><td> 93</td><td> 87</td><td> 95</td><td> 88</td><td> 96</td><td> 89</td><td> 97</td>

<td rowspan="2">Chains heavy Ab6</td><td> 86</td><td> 94</td><td> 90</td><td> 98</td><td> 91</td><td> 99</td><td> 92</td><td> 100</td>

<td rowspan="4">Chains light of Ab7</td><td> 101</td><td> 109</td><td> 103</td><td> 111</td><td> 104</td><td> 112</td><td> 105</td><td> 113</td>

<td rowspan="6">Chains heavy Ab7</td><td> 102</td><td> 110</td><td> 106</td><td> 114</td><td> 107</td><td> 115</td><td> 108</td><td> 116</td>

240

IMPI

<td rowspan="2">Light chain from Ab8</td><td rowspan="2"> 122</td><td rowspan="2"> 130</td><td rowspan="2"> 124</td><td rowspan="2"> 132</td><td></td><td>INDUS1 and</td><td>TO THE</td><td rowspan="2"> 134</td>

<td>Heavy chain from Ab8</td><td> 123</td><td> 131</td><td> 127</td><td> 135</td><td> 128</td><td> 136</td><td> 129</td><td> 137</td>

<td>Light chain from Ab9</td><td> 138</td><td> 146</td><td> 140</td><td> 148</td><td> 141</td><td> 149</td><td> 142</td><td> 150</td>

<td>Heavy chain from Ab9</td><td> 139</td><td> 147</td><td> 143</td><td> 151</td><td> 144</td><td> 152</td><td> 145</td><td> 153</td>

<td>Light chain from AblO</td><td> 154</td><td> 162</td><td> 156</td><td> 164</td><td> 157</td><td> 165</td><td> 158</td><td> 166</td>

<td>Heavy chain from AblO</td><td> 155</td><td> 163</td><td> 159</td><td> 167</td><td> 160</td><td> 168</td><td> 161</td><td> 169</td>

<td>Light chain from Abll</td><td> 170</td><td> 178</td><td> 172</td><td> 180</td><td> 173</td><td> 181</td><td> 174</td><td> 182</td>

<td>Heavy chain from Abll</td><td> 171</td><td> 179</td><td> 175</td><td> 183</td><td> 176</td><td> 184</td><td> 177</td><td> 185</td>

<td>Light chain by Abl2</td><td> 186</td><td> 194</td><td> 188</td><td> 196</td><td> 189</td><td> 197</td><td> 190</td><td> 198</td>

<td>Heavy chain by Abl2</td><td> 187</td><td> 195</td><td> 191</td><td> 199</td><td> 192</td><td> 200</td><td> 193</td><td> 201</td>

<td>Light chain by Abl3</td><td> 202</td><td> 210</td><td> 204</td><td> 212</td><td> 205</td><td> 213</td><td> 206</td><td> 214</td>

241 ίΐ ο I f'-EA 1

KFXICANO INSTITUTE

<td></td><td></td><td></td><td></td><td></td><td></td><td>I</td><td>IDuSTillAL</td><td></td>

<td>Heavy chain</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>Light chain from Afc> 14</td><td> 218</td><td> 226</td><td> 220</td><td> 228</td><td> 221</td><td> 229</td><td> 222</td><td> 230</td>

<td>Heavy chain by Abl4</td><td> 219</td><td> 227</td><td> 223</td><td> 231</td><td> 224</td><td> 232</td><td> 225</td><td> 233</td>

<td>Light chain by Abl5</td><td> 234</td><td> 242</td><td> 236</td><td> 244</td><td> 237</td><td> 245</td><td> 238</td><td> 246</td>

<td>Heavy chain from Afc> 15</td><td> 235</td><td> 243</td><td> 239</td><td> 247</td><td> 240</td><td> 248</td><td> 241</td><td> 249</td>

<td>Light chain from Abl6</td><td> 250</td><td> 258</td><td> 252</td><td> 260</td><td> 253</td><td> 261</td><td> 254</td><td> 262</td>

<td>Heavy chain from Abl6</td><td> 251</td><td> 259</td><td> 255</td><td> 263</td><td> 256</td><td> 264</td><td> 257</td><td> 265</td>

<td>Light chain by Abl7</td><td> 266</td><td> 274</td><td> 268</td><td> 276</td><td> 269</td><td> 277</td><td> 270</td><td> 278</td>

<td>Heavy chain by Abl7</td><td> 267</td><td> 275</td><td> 271</td><td> 279</td><td> 272</td><td> 280</td><td> 273</td><td> 281</td>

<td>Light chain by Abl8</td><td> 282</td><td> 290</td><td> 284</td><td> 292</td><td> 285</td><td> 293</td><td> 286</td><td> 294</td>

<td>Heavy chain by Abl8</td><td> 283</td><td> 291</td><td> 287</td><td> 295</td><td> 288</td><td> 296</td><td> 289</td><td> 297</td>

242

IMPI

<td rowspan="2">Light chain by Abl9</td><td rowspan="2"> 298</td><td rowspan="2"> 306</td><td rowspan="2"> 300</td><td rowspan="2"> 308</td><td></td><td>INDUS1</td><td>JAL) UAL</td><td rowspan="2"> 310</td>

<td>Heavy chain by Abl9</td><td> 299</td><td> 307</td><td> 303</td><td> 311</td><td> 304</td><td> 312</td><td> 305</td><td> 313</td>

<td>Light chain from Ab20</td><td> 314</td><td> 322</td><td> 316</td><td> 324</td><td> 317</td><td> 325</td><td> 318</td><td> 326</td>

<td>Heavy chain from Ab20</td><td> 315</td><td> 323</td><td> 319</td><td> 327</td><td> 320</td><td> 328</td><td> 321</td><td> 329</td>

<td>Light chain from Ab21</td><td> 330</td><td> 338</td><td> 332</td><td> 340</td><td> 333</td><td> 341</td><td> 334</td><td> 342</td>

<td>Heavy chain from Ab21</td><td> 331</td><td> 339</td><td> 335</td><td> 343</td><td> 336</td><td> 344</td><td> 337</td><td> 345</td>

<td>Light chain from Ab22</td><td> 346</td><td> 354</td><td> 348</td><td> 356</td><td> 349</td><td> 357</td><td> 350</td><td> 358</td>

<td>Heavy chain from Ab22</td><td> 347</td><td> 355</td><td> 351</td><td> 359</td><td> 352</td><td> 360</td><td> 353</td><td> 361</td>

<td>Light chain from Ab23</td><td> 362</td><td> 370</td><td> 364</td><td> 372</td><td> 365</td><td> 373</td><td> 366</td><td> 374</td>

<td>Heavy chain from Al> 23</td><td> 363</td><td> 371</td><td> 367</td><td> 375</td><td> 368</td><td> 376</td><td> 369</td><td> 377</td>

<td>Light chain by Ab24</td><td> 378</td><td> 386</td><td> 380</td><td> 388</td><td> 381</td><td> 389</td><td> 382</td><td> 390</td>

243

244

IMPI

<td rowspan="2">Light chain from Ab30</td><td rowspan="2"> 474</td><td rowspan="2"> 482</td><td rowspan="2"> 476</td><td rowspan="2"> 484</td><td></td><td>OF THE PROF indu;</td><td>AGE CV TR1AL</td><td rowspan="2"> ~*486~</td>

<td>Heavy chain from Ab30</td><td> 475</td><td> 483</td><td> 479</td><td> 487</td><td> 480</td><td> 488</td><td> 481</td><td> 489</td>

<td>Light chain from Ab31</td><td> 490</td><td> 498</td><td> 492</td><td> 500</td><td> 493</td><td> 501</td><td> 494</td><td> 502</td>

<td>Heavy chain from Ab31</td><td> 491</td><td> 499</td><td> 495</td><td> 503</td><td> 496</td><td> 504</td><td> 497</td><td> 505</td>

<td>Light chain by Ab32</td><td> 506</td><td> 514</td><td> 508</td><td> 516</td><td> 509</td><td> 517</td><td> 510</td><td> 518</td>

<td>Heavy chain by Ab32</td><td> 507</td><td> 515</td><td> 511</td><td> 519</td><td> 512</td><td> 520</td><td> 513</td><td> 521</td>

<td>Light chain from Ab33</td><td> 522</td><td> 530</td><td> 524</td><td> 532</td><td> 525</td><td> 533</td><td> 526</td><td> 534</td>

<td>Heavy chain from Ab33</td><td> 523</td><td> 531</td><td> 527</td><td> 535</td><td> 528</td><td> 536</td><td> 529</td><td> 537</td>

<td>Light chain from Ab34</td><td> 538</td><td> 546</td><td> 540</td><td> 548</td><td> 541</td><td> 549</td><td> 542</td><td> 550</td>

<td>Heavy chain from Ab34</td><td> 539</td><td> 547</td><td> 543</td><td> 551</td><td> 544</td><td> 552</td><td> 545</td><td> 553</td>

<td>Light chain from Ab35</td><td> 554</td><td> 562</td><td> 556</td><td> 564</td><td> 557</td><td> 565</td><td> 558</td><td> 566</td>

245

IMPI

<td rowspan="2">Heavy chain from Ab35</td><td rowspan="2"> 555</td><td rowspan="2"> 563</td><td rowspan="2"> 559</td><td rowspan="2"> 567</td><td></td><td>OF THE PROi INDI</td><td>PIETY STR1AL</td><td></td>

<td>Light chain by Ab36</td><td> 570</td><td> 578</td><td> 572</td><td> 580</td><td> 573</td><td> 581</td><td> 574</td><td> 582</td>

<td>Heavy chain by Ab36</td><td> 571</td><td> 579</td><td> 575</td><td> 583</td><td> 576</td><td> 584</td><td> 577</td><td> 585</td>

* Examples of heavy and light chain sequence variant forms are shown on separate lines.

PRT .: Polypeptide sequence

Nuc .: Examples of coding sequence.

For reference, sequence identifiers other than those included in Table 1 are summarized in Table

the

Table present

2. Summary of request sequence identifiers.

<td>SEQ ID</td><td>Description</td>

<td> 1</td><td>Human IL-6</td>

<td> 586</td><td>chain polypeptide sequence slight constant kappa</td>

<td> 587</td><td>chain polynucleotide sequence slight constant kappa</td>

<td> 588</td><td>chain polypeptide sequence</td>

246

MEXICAN INSTITUTE

<td></td><td>INDUSTRIAL gamma-1 constant weighing</td>

<td> 589</td><td>chain polynucleotide sequence gamma-1 constant weighing</td>

<td> 590-646</td><td>Human IL-6 peptides (see FIG. 12 and Example 14)</td>

<td> 719</td><td>Chain polypeptide sequence constant gamma-1 weight (differs from SEQ ID NO: 518 in two positions)</td>

<td> 726</td><td>Protein polypeptide sequence C reactive</td>

<td> 727</td><td>IL-6 alpha receptor</td>

<td> 728</td><td>IL-6 beta / gp receptor! 30</td>

Said antibody fragments may be present in one or more of the following non-restrictive forms: Fab, Fab ',

F (ab ') 2z Fv and Fv single chain antibody forms. In a preferred embodiment, the anti-IL-6 antibodies described herein further comprise the kappa constant light chain sequence comprising the sequence set forth below:

VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ

DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:

586) .

In another preferred embodiment, the anti-IL-6 antibodies described herein further comprise the sequence

247

MEXICAN INSTITUTE OF THE rrX'PlFOAD IN.O'JS't'ÍUAL of gamma-1 constant heavy chain polypeptide comprising one of the sequences established below:

ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL

QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGG

PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTY

RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQV

SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS

VMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 588) and

ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL

QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGG

PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTY

RWS VLT VLHQDWLNGKE YKCKVSNKAL PAPIΕΚΤIS KAKGQ PRE PQVYTLP PS RDELTKNQV

SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS

VMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 719).

The modalities of the antibodies described herein may include a leader sequence, such as, a leader

Rabbit Ig, an albumin prepeptide, a yeast mating factor pre or pro secretion leader sequence (such as P. pastoris or Saccharomyces cerevisiae or alpha factor) or a human HAS leader. The examples of leader sequences are shown misaligned with respect to FRl at the N-terminus of the polypeptides shown in Figures 36A and 37A as follows: mouse Ig leader sequences of SEQ ID NOs: 2 and 660 (MD... ) and SEQ ID NOs: 3 and 661 (ME ...) and an albumin prepeptide

248

ΙΜΡΐ

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

in SEQ ID NOs: 706 and 708, which facilitates its secretion. Other leader sequences known in the art can also be used to confer desired properties, such as improved secretion, stability or half-life, etc., alone or in combination with each other, in the heavy and / or light chains, which can optionally be cleaved prior to administration to a subject. For example, a polypeptide can be expressed in a cell-free or cell expression system that also expresses or includes (or is modified to express or include) a protease, eg, a membrane-linked signal peptidase that cleaves a sequence Leader.

In another embodiment, the invention contemplates an antibody

<td>anti-IL-6</td><td>isolated that</td><td>understands</td><td>a sequence of</td><td>polypeptide of</td>

<td colspan="2">VH which includes:</td><td>SEQ ID NO:</td><td> 3, 18, 19, 22,</td><td> 38, 54, 70, 86,</td>

<td colspan="5">708 and also comprises a V polypeptide sequenceL than</td>

<td>understands:</td><td>the SEQ ID</td><td>NO: 2, 20</td><td> , 21, 37, 53, 69</td><td> , 85, 101, 119,</td>

<td> 679, 683,</td><td colspan="3">687, 693, 699, 702, 706, or 709 or a</td><td>variant of the</td>

same where one or more of the residues flanking residues

249

IMPI

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

FR) in. said polypeptide of VH or VL it was replaced by another amino acid residue resulting in an anti-IL-6 antibody that specifically binds IL-6. The invention contemplates humanized and chimeric forms of these antibodies. Chimeric antibodies can include an Fe derived from the IgGl, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7 constant regions,

IgG8, IgG9, IgGlO, IgGll, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17, IgG18 and IgG19 and in particular a variable heavy and light chain constant region such as that contained in SEQ ID NO: 588 and SEQ ID NO : 586.

In one embodiment of the invention, the V antibodies or polypeptidesH or VL they originate or are selected from one or more rabbit B lymphocyte populations prior to the start of the humanization process named herein.

In another embodiment of the invention, anti-IL-6 antibodies and fragments thereof possess specificity for primate homologs of human IL-6 protein. Non-limiting examples of primate homologs of human IL-6 protein are IL-6 obtained from Macaca fascicularis (also known as macaque monkey) and Rhesus monkey. In another embodiment of the invention, anti-IL-6 antibodies and fragments thereof inhibit the association of IL-6 with IL6R, and / or the production of IL-6 / IL-6R / gpl30 complexes and / or the production of IL-6 / IL-6R / gpl30 multimers and / or antagonize the biological effects of one or more of the aforementioned.

250

As stated above, antibodies and fragments thereof can be post-translationally modified to add effector residues such as chemical bonds, detectable residues such as, for example, fluorescent dyes, enzymes, substrates, bioluminescent materials, radioactive materials, and chemiluminescent residues or residues. functional such as, for example, streptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, and radioactive materials.

As for detectable residues, additional examples of enzymes include, but are not limited to, horseradish peroxidase, acetylcholinesterase, alkaline phosphatase, beta-galactosidase, and luciferase. Additional examples of fluorescent materials include, but are not limited to, rhodamine, fluorescein, fluorescein isothiocyanate, umbelliferone, dichlorotriazinylamine, phycoerythrin, and dansyl chloride. Additional examples of chemiluminescent residues include, but are not limited to, luminol. Additional examples of bioluminescent materials include, but are not limited to, luciferin and aequorin. Additional examples of radioactive materials include, but are not limited to, Iodine 125 '125-1

35r

Carbon-14 (14C), Sulfur 35 (J0S), Tritium (JH) and Phosphorus 32 (32P) ·

As for functional residues, examples of cytotoxic agents include, but are not limited to, methotrexate,

251 '7;

IMPIOS

VLXICAN INSTITUTE ú £ LA? ÍO?. '£ DAi>

INDUSTRIAL ___ aminopterin, 6-mercaptopurma, 6-thioguanma, cytarabine, 5fluorouracil dacarbazine, alkylating agents such as mechlorethamine, thioepa chlorambucil, melphalan, carmustine, mitomycin C, lomustine (1) , dibromomanitol, streptozotocin, mitomycin C, cis-dichlorodiamine platinum (II) (DDP) cisplatin and carboplatin (paraplatin); the

<td>anthracyclines</td><td></td><td>include</td><td>daunorubicin</td><td>(formerly</td>

<td>daunomycin),</td><td colspan="2">doxorubicin</td><td>(adriamycin),</td><td>detorubicin,</td>

<td>carminomycin,</td><td colspan="2">idarubicin,</td><td>epirubicin,</td><td>mitoxantrone and</td>

<td>bisantrene;</td><td>the</td><td colspan="2">antibiotics include</td><td>dactinomycin</td>

<td>(actinomycin</td><td>D),</td><td>bleomycin,</td><td>calicheamicin</td><td>, mithramycin and</td>

<td colspan="2">anthramycin (AMC)</td><td>and agents</td><td>antimitotics</td><td>such as</td>

<td>alkaloids</td><td>the</td><td colspan="3">vinca, vincristine and vinblastine. Others</td>

Cytotoxic agents include paclitaxel (taxol), ricin, pseudomonas exotoxin, gemcitabine, cytochalasin B, gramicidin

D, ethidium bromide, emetine, etoposide, tenoposide, colchicine, dihydroxy anthracyn dione, 1-dihydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, procarbazine, hydroxyurea, asparaginase, corticosteroids, - ( DDD)), interferons and mixtures of these cytotoxic agents.

Additional cytotoxic agents include, but are not limited to, chemotherapeutic agents such as carboplatin, cisplatin, paclitaxel, gemcitabine, calicheamycin,

252

doxorubicin, 5-fluorouracil, mitomycin C, actinomycin D, cyclophosphamide, vincristine, bleomycin, VEGF antagonists, EGFR antagonists, platins, taxols, irinotecan, 5fluorouracil, gemcitabine, leucovorin, spheroids, cyclophosphamide, motelophosphamide, cyclophosphamide, porcine cyclophosphamide (eg, vinblastine, vincristine, vindesine, and vinorelbine), mustins, tyrosine kinase inhibitors, radiation therapy, sex hormone antagonists, selective androgen receptor modulators, selective estrogen receptor modulators, PDGF antagonists, TNF antagonists, IL1 antagonists, interleukins (eg IL-12 or IL-2), IL-12R antagonists, toxin-conjugated monoclonal antibodies, antigen-specific monoclonal antibodies tumors, Erbitux ™, Avastin ™, Pertuzumab, anti-CD20 antibodies, Rituxan®, ocrelizumab, ofatumumab, DXL625, Herceptin® or any combination thereof. Toxic plant and bacterial enzymes such as ricin, diphtheria toxin, and Pseudomonas toxin can conjugate with humanized antibodies or binding fragments thereof to generate reagents for specific destruction of a cell type (Youle, et al., Proc. Nat 'l Acad. Sci. USA 77: 5483 (1980); Gilliland, et al., Proc. Nat'l Acad. Sci. USA 77: 4539 (1980); Krolick, et al., Proc. Nat'l Acad. Sci. USA 77: 5419 (1980)).

Other cytotoxic agents include cytotoxic ribonucleases as described by Goldenberg in the Patent

253 No. 6,653,104

The embodiments of the invention also relate to radioimmunoconjugates where an alpha or beta particle emitting radionuclide is stably coupled to the antibody, or binding fragments thereof, with or without the use of a complexing agent.

Such radionuclides include beta emitters such as Phosphorus32 (32P), Scandium-47 (47Sc), Copper-67 (67Cu), Gallium-67 ( 67Ga),

Yttrium-88 (88Y), Yttrium-90 (90Y), Iodine-125 (125I), Iodine-131 (131I), Samario-153 (153Sm), Lutetium-177 (177Lu), Renio-186 (186Re) or Rhenium-188 (188Re), and alpha emitters such as Astatin-211 (211At), Lead-212 (212Pb), Bismuth-212 (212Bi) or -213 (213Bi) o

Actinium-225 (225Ac).

Methods are known in the art for conjugation of an antibody or a binding fragment thereof to a detectable moiety and the like, such as, for example, the methods described by Hunter et al, Nature 144: 945 (1962); David et al,

Biochemistry 13: 1014 (1974); Pain et al, J. Immunol. Meth.

40: 219 (1981) and Nygren, J., Histochem. and Cytochem. 30: 407 (1982).

The modalities described herein further include variants and equivalents substantially homologous to antibodies, antibody fragments, diabodies, SMIPs, camelid antibodies, nanoantibodies, IgNARs, polypeptides, variable regions and CDRs set forth herein. These may contain, for example, mutations of

254

conservative substitution (that is, the substitution of one or more amino acids by means of similar amino acids). For example, conservative substitution refers to the substitution of one amino acid for another within the same general class, for example, an acidic amino acid for another acidic amino acid, a basic amino acid for another basic amino acid, or a neutral amino acid for another neutral amino acid. What is intended by a conservative substitution of an amino acid is well known in the art.

In another embodiment, the invention contemplates polypeptide sequences that possess at least 90% or more sequence homology to any one or more of the antibody fragment, variable region, and CDR polypeptide sequences set forth herein. More preferably, the invention contemplates polypeptide sequences that possess at least 95% or more sequence homology, more preferably, at least 98% or more sequence homology, and even more preferably at least 99% or more sequence homology. sequence with any one or more of the antibody fragment, variable region and CDR polypeptide sequences set forth herein. Methods for determining homology between nucleic acid and amino acid sequences are well known to those skilled in the art.

In another embodiment, the invention further contemplates the aforementioned fragment polypeptide homologs

255

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MEXICAN INSTITUTE OF INDUSTRIAL INSTITUTE of antibody, variable regions and CDRs established herein that additionally possess anti-IL-6 activity.

Non-limiting examples of anti-IL-6 activity are set forth herein, for example, under the heading "Anti-IL6 Activity" below.

In another embodiment, the invention further contemplates the creation and use of anti-idiotypic antibodies that bind to any of the preceding sequences. In an exemplary embodiment, this anti-idiotypic antibody can be administered to a subject who received an anti-IL-6 antibody to modulate, reduce, or neutralize the effect of the anti-IL-6 antibody. Such anti-idiotypic antibodies may also be useful in the treatment of autoimmune diseases characterized by the presence of anti-IL-6 antibodies. A further example of use of such anti-idotypic antibodies is the detection of anti-IL-6 antibodies of the present invention, for example, to control the levels of anti-IL-6 antibodies present in the blood or other body fluids of a subject.

The present invention also contemplates anti-IL-6 antibodies comprising any of the polypeptide or polynucleotide sequences described herein that replace any of the other polynucleotide sequences described herein. By way of example and not exhaustively, the present invention contemplates antibodies of

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MEXICAN INSTITUTE, n ,. . ,,. OF PROPERTY comprising the combination of any one variable-light chain and one variable-heavy chain · do SGJÍ¿tAS ^ -an ^ - ^ The present and additionally contemplates antibodies resulting from the substitution of any of the other CDR sequences described herein by any of the CDR sequences described herein.

Examples of additional embodiments of the invention

In another embodiment, the invention contemplates one or more anti-IL-6 antibodies or antibody fragment that can specifically bind to the same linear or conformational epitope (s) and / or compete to bind to the same linear or conformational epitope (s) in an intact human IL-6 polypeptide or fragment thereof as an anti-IL-6 antibody comprising Abl, Ab2, Ab3 , Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, AblO, Abll, Abl2, Abl3, Abl4, Abl5, Abl6,

Abl7, Abl8, Abl9, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25, Ab26,

Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, or Ab36 and humanized, single-chain chimeric antibodies and fragments thereof (containing one or more CDRs of the aforementioned antibodies) that bind specifically IL-6, which is preferably aglycosylated. In a preferred embodiment, the anti-IL-6 antibody or fragment thereof can specifically bind to the same linear or conformational epitope (s) and / or compete to bind to the same (s)

257

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL linear or conformational epitope (s) in an intact human IL-6 polypeptide or a fragment thereof such as Abl or an antibody comprising Abl CDRs.

In another embodiment of the invention, anti-IL6 antibody that specifically binds to the same linear or conformational epitopes on an intact IL-6 polypeptide or a fragment thereof that is specifically bound by Abl, binds to one / more epitopes / s of IL-6 determined by epitopic mapping using overlapping linear polypeptide fragments spanning the entire length of native human IL-6 polypeptide. In one embodiment of the invention, the IL-6 epitope comprises or, alternatively, consists of one or more residues comprised of fragments of IL-6 selected from those comprising respectively amino acid residues 37-51, the amino acid residues 70-84, amino acid residues 169-183, amino acid residues 31-45 and / or amino acid residues 58-72.

The invention is also directed to an anti-IL-6 antibody that binds to the same IL-6 epitope and / or competes with an anti-IL-6 antibody to bind IL-6 as an antibody or antibody fragment described in herein, including but not limited to, an anti-IL-6 antibody selected from

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MEXICAN INSTITUTE OF INDUSTRIAL AGE

Ab32, Ab33, Ab34, Ab35, or Ab36 and humanized, single-chain chimeric antibodies and fragments thereof (containing one or more CDRs of the aforementioned antibodies) that specifically bind to IL-6, which are preferably aglycosylated.

In another embodiment, the invention is also directed to an isolated antibody or anti-IL-6 antibody fragment comprising one or more of the CDRs contained in the sequences of

<td>V polypeptideH which include:</td><td>I KNOW THAT</td><td>ID NO:</td><td> 3, 18, 19, 22,</td>

<td colspan="2">691, 692, 704, or 708 and / or one or more of the</td><td colspan="2">CDR contained in the</td>

<td>V polypeptide sequenceL than</td><td colspan="2">understands:</td><td> 2, 20, 21, 37,</td>

<td colspan="3">the VH and VL sequences represented in the</td><td>alignments of</td>

antibodies included in Figures 34-37 of the present application.

In one embodiment of the invention, the anti-IL-6 antibody referred to in the previous two paragraphs comprises at least 2 regions determining

259

IMPI

• · · 's “-Λίν.η., ν,

PROPERTY V 'INDUSTRIAL * Complementarity (CDR) in each variable light and variable heavy region that are identical to those contained in an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4, Ab5,

Ab6, Ab7, Ab8, Ab9, AblO, Abll, Abl2, Abl3, Abl4, Abl5, Abl6,

Abl7, Abl8, Abl9, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25, Ab26,

Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, or Ab36 and single chain humanized chimeric antibodies and fragments thereof (containing one or more CDRs of the aforementioned antibodies) that bind specifically IL-6, which is preferably aglycosylated.

In a preferred embodiment, the anti-IL-6 antibody referred to above comprises at least 2 complementarity determining regions (CDRs) in each of the variable light and variable heavy regions that are identical to those contained in Abl. In another embodiment, all the CDRs of the anti-IL-6 antibody referenced above are identical to the CDRs contained in an anti-IL-6 antibody comprising Abl, Ab2, Ab3, Ab4, Ab5,

Ab6, Ab7, Ab8, Ab9, AblO, Abll, Abl2, Abl3, Abl4, Abl5, Abl6,

Abl7, Abl8, Abl9, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25, Ab26,

Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, or Ab36 or humanized, single-chain chimeric antibodies and fragments thereof (containing one or more CDRs of the aforementioned antibodies) that bind specifically IL-6, which is preferably aglycosylated. In a modality

260

IMPIOS

MEXICAN INSTITUTE \ 'Λ », Λ

PILA W.'WEL'AO VVn.—

INDUSTRIAL preferred of the invention, all the CDRs of the antiIL-6 antibody indicated above are identical to the CDRs contained in Abl, for example, an antibody comprising the VH and VL sequences comprised in SEQ ID NO: 657 and SEQ ID NO : 709 respectively.

The invention further contemplates that the anti-IL-6 antibody or antibodies indicated above are akylated, containing an Ec region that has been modified to alter the effector function, half-life, proteolysis and / or glycolysation, are human, humanized, single-stranded or chimeric and are a humanized antibody that comes from a rabbit anti-IL-6 antibody (parent).

Examples of constant regions that provide the production of aglycosylated antibodies in Pichia are included in the

SEQ ID NO: 588 and SEQ ID NO: 586 which are respectively encoded by the nucleic acid sequences in SEQ ID NO: 589 and SEQ

ID NO: 587.

The invention further contemplates one or more anti-IL-6 antibodies where the flanking regions (FR) in the variable light region and the variable heavy regions of said antibody are respectively unmodified human FR or that have been modified by substituting at most 2 or 3 human FR residues in the light or heavy chain region variable with the corresponding FR residues of the parent rabbit antibody and where said human FR

261

IMP

I

come from human variable weight and light chain antibody sequences that have been 3616001011530 ^^ 31 ^^ 1113 library of human germline antibody sequences, based on their high level of homology to the corresponding rabbit variable light or heavy chain regions with relative to other human germ antibody sequences contained in the library.

In one embodiment of the invention, the anti-IL-6 antibody or fragment thereof can specifically bind to human cells expressing IL-6 and / or to circulating soluble IL-6 molecules, including IL-6 expressed in or by human cells in a patient suffering from a disease associated with cells expressing IL-6.

In another modality, the disease is selected from general fatigue, exercise-induced fatigue, cancer-related fatigue, inflammatory disease-related fatigue, chronic fatigue syndrome, fibromyalgia, cancerous cachexia, cardiac cachexia, respiratory cachexia, renal cachexia, cachexia age-related rheumatoid arthritis, systemic lupus erythematosus (SLE), systemic juvenile idiopathic arthritis, psoriasis, psoriatic arthropathy, ankylosing spondylitis, inflammatory bowel disease (IBD), polymyalgia rheumatica, giant cell arteritis, autoimmune vasculitis, chronic graft versus host disease (GVHD), Sjogren's syndrome, Still's disease of

262

IMPI

adult onset, rheumatoid arthritis, systemic juvenile idiopathic arthritis, osteoarthritis, osteoporosis, Paget's disease of bone, osteoarthritis, multiple myeloma, Hodgkin lymphoma, non-Hodgkin lymphoma, prostate cancer, leukemia, renal cell cancer, Castleman's disease multicenter, ovarian cancer, drug tolerance in cancer chemotherapy, toxicity of cancer chemotherapy, ischemic heart disease, atherosclerosis, obesity, diabetes, asthma, multiple sclerosis, Alzheimer's disease, cerebrovascular disease, fever, acute phase response, allergies, anemia, anemia of inflammation (anemia of chronic disease), hypertension, depression, depression associated with chronic disease, thrombosis, thrombocytosis, acute heart failure , metabolic syndrome, miscarriages, obesity, chronic prostatitis, glomerulonephritis, pelvic inflammatory disease, reperfusion injury, transplant rejection, graft versus host disease (GVHD), avian influenza, smallpox, pandemic influenza, adult respiratory distress syndrome (ARDS), severe acute respiratory syndrome (SARS), sepsis, and systemic inflammatory response syndrome (SIRS).

In a preferred embodiment, the disease is selected from cancer, inflammatory disorder, viral disorder, or autoimmune disorder. In a particularly preferred embodiment, the disease is arthritis, cachexia, and wasting syndrome.

The invention further contemplates antibodies or

263

anti-IL-6 antibody fragments directly or indirectly linked to a detectable label or a therapeutic agent.

The invention also contemplates one or more nucleic acid sequences that result in expression of an antibody or antibody fragment as set forth above, including those that comprise, or alternatively consist of, preferred yeast or human codons. The invention also contemplates vectors (including viral plasmids or recombinant vectors) comprising said nucleic acid sequence (s). The invention also contemplates host cells or recombinant host cells that express at least one of the above established antibodies, including mammalian, yeast, bacterial, and insect cells. In a preferred embodiment, the host cell is a yeast cell. In a further preferred embodiment, the yeast cell is a diploid yeast cell. In an even more preferred embodiment, the yeast cell is a Pichia cell.

The invention also contemplates a method of treatment comprising administering to a patient suffering from a disease or condition associated with cells expressing IL-6, a therapeutically effective amount of at least one IL-6 antibody or antibody fragment. The diseases that can be treated are presented in the non-exhaustive list established above. In a preferred embodiment, the

264

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY disease is selected from cancer, autoimmune disease or inflammatory condition. In a particularly preferred embodiment, the disease is cancer or viral infection. In another embodiment, treatment additionally includes administration of another agent or therapeutic regimen selected from chemotherapy, radiation therapy, cytokine administration, or gene therapy.

The invention further contemplates an in vivo imaging method that detects the presence of cells expressing IL-6 which comprises administering a diagnostic effective amount of at least one anti-IL-6 antibody. In one embodiment, such administration further includes administration of a radionuclide or fluorophore that facilitates detection of the antibody at IL-6 expressing disease sites. In another embodiment of the invention, the in vivo imaging method is used to detect tumors or metastases that express IL-6 or is used to detect the presence of autoimmune disorder sites associated with cells that express IL-6. In a further embodiment, the results of such an in vivo imaging method are used to facilitate the design of an appropriate therapeutic regimen that includes therapeutic regimens including radiation therapy, chemotherapy, or a combination thereof.

Polynucleotides Encoding Anti265 Antibody Polypeptides

IMPI

IL-6

The invention is further directed to polynucleotides encoding polypeptides of antibodies that possess IL-6 binding specificity. In an embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the variable light chain polypeptide sequence of SEQ ID NO: 2:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAA

CCAGGGCAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCG

CGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCC

GATGCTGCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCTTTCGGC

GGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCA

TCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTT (SEQ

ID NO: 10)

In another embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID

NO: 3:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACAGCCTCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGCCAGGCTCCAGGGAAG

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IMPI

GGGCTGGAATGGATCGGAATCATTTATGGTAGTGATGAAACGGCCTACGCGACCTGGGCGATA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCC

GCGGACACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAAC

TTGTGGGGCCAAGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTC

CCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAG

G (SEQ ID NO: 11).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 12; SEQ ID NO: 13 and SEQ ID NO: 14 which correspond to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 2.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 15; SEQ ID NO: 16 and SEQ ID NO: 17 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 3.

The invention also contemplates sequences of

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INSTITUTO MEXiCANO V Dt LA INDUSTRIAL PROPERTY polynucleotides that include one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 10 encoding the variable light chain region of SEQ ID NO: 2, the polynucleotide of SEQ ID NO: eleven encoding the variable heavy chain region of SEQ ID NO: 3; polynucleotides encoding complementarity determining regions (SEQ ID NO: 12; SEQ ID NO: 13 and SEQ ID NO: 14) of the variable light chain region of SEQ ID NO: 10; polynucleotides that encode complementarity determining regions (SEQ ID NO: 15; SEQ ID NO: 16 and SEQ ID NO: 17) of the variable heavy chain region of SEQ ID NO: 11 and polynucleotides encoding the variable light and heavy chain sequences in SEQ ID NO: 657 and SEQ ID NO: 709 respectively, eg, the nucleic acid sequences in SEQ ID NO: 700 and SEQ ID NO: 723 and fragments or variants thereof, eg, based on codon degeneration. These nucleic acid sequences encoding variable heavy and light chain sequences can be expressed alone or in combination and these sequences are preferably fused to variable constant sequences.

268

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INSTITUTO MEXICANO DE LA ríOPIEDAU INDUSTRIAL suitable, for example, those in SEQ ID NO: 589 and SEQ ID NO:

587.

The invention is further directed to poiinucleotides encoding polypeptides of antibodies that possess IL-6 binding specificity. In one embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following poiinucleotide sequence encoding the variable light chain polypeptide sequence of SEQ ID NO: 21:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACA

GTCACCATCAATTGCCAGGCCAGTGAGACCATTTACAGTTGGTTATCCTGGTATCAGCAGAAG

CCAGGGCAGCCTCCCAAGCTCCTGATCTACCAGGCATCCGATCTGGCATCTGGGGTCCCATCG

CGATTCAGCGGOAGTGGGGCTGGGACAGAGTACACTCTCACCATCAGCGGCGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTCAACAGGGTTATAGTGGTAGTAATGTTGATAATGTTTTCGGC

GGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCA

TCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCC

AGAGAGGCCAAAG (SEQ ID NO: 29)

In another embodiment of the invention, the poiinucleotides of the invention comprise or, alternatively, consist of the following poiinucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID

NO: 22:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTTACC

269

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

TGCACAGCCTCTGGATTCTCCCTCAATGACCATGCAATGGGCTGGGTCCGCCAGGCTCCAGGG

AAGGGGCTGGAATACATCGGATTCATTAATAGTGGTGGTAGCGCACGCTACGCGAGCTGGGCA

GAAGGCCGATTCACCATCTCCAGAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACA

ACCGAGGACACGGCCACCTATTTCTGTGTCAGAGGGGGTGCTGTTTGGAGTATTCATAGTTTT

GATCCCTGGGGCCCAGGGACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTC

TTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTC

AAG (SEQ ID NO: 30).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 31; SEQ ID NO: 32 and SEQ ID NO: 33 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 21.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 34; SEQ ID NO: 35 and SEQ ID NO: 36 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 22.

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IMPI

The invention also contemplates polynucleotide sequences that include one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 29 encoding the variable light chain region of SEQ ID NO: 21, the polynucleotide of SEQ ID NO: 30 encoding the variable heavy chain region of SEQ ID NO: 22; polynucleotides that encode the complementarity determining regions (SEQ ID NO: 31; SEQ ID NO: 32 and SEQ ID NO: 33) of the region of

<td>light chain</td><td>variable</td><td>of</td><td>the SEQ ID</td><td>NOT:</td><td> 29;</td><td>and</td><td>the</td>

<td>polynucleotides</td><td colspan="2">that encode</td><td>the regions</td><td colspan="3">determinants</td><td>of</td>

<td colspan="2">complementarity (SEQ ID</td><td>NOT:</td><td>3. 4; SEQ ID NO:</td><td> 35</td><td>and SEQ</td><td>ID</td><td>NOT:</td>

<td>36) from the region</td><td>chain</td><td colspan="2">variable weighing</td><td>the</td><td>SEQ ID</td><td>NOT:</td><td> 30.</td>

The invention is further directed to polynucleotides encoding polypeptides of antibodies that possess IL-6 binding specificity. In one embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the polypeptide sequence of

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INDUSTRIAL

variable light chain of SEQ ID NO: 37:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACA

GTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATGACAACAACTACTTATCCTGGTTTCAG

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTC

CCATCGCGGTTCGTGGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACAGACGTGCAG

TGTGACGATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATAATGCCTTC

GGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCG

CCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCT (SEQ ID NO: 45)

NO: 38:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACCCCTGGGACACCCCTGACACTCACCTGC

ACAGCCTCTGGATTCTCCCTCAGTGTCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGATTCATTACAATGAGTGATAATATAAATTACGCGAGCTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTACAATGGGTCGGTTGGAT

CTCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTC

CCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAG

G (SEQ ID NO: 46).

In a further embodiment of the invention, the

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY polynucleotides encoding fragments of the antibody possessing specificity for binding to IL-6 comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 47; SEQ ID NO: 48 and SEQ ID NO:

corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 37.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 50; SEQ ID NO: 51 and SEQ ID NO: 52 which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td>heavy chain</td><td>variable of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

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INDUSTRIAL

Antibody: the polynucleotide of SEQ ID NO: 45 encoding the variable light chain region of SEQ ID NO: 37, the polynucleotide of SEQ ID NO: 46 encoding the variable heavy chain region of SEQ ID NO: 38; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49) of the variable light chain region of SEQ ID NO: 37; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 50; SEQ ID NO: 51 and SEQ ID NO: 52) of the variable heavy chain region of SEQ ID NO: 38.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCATATGTGACCCTGTGCTGACCCAGACTCCATCTCCCGTATCTGCACCTGTGGGAGGCACA

GTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATGAGAACAACTATTTATCCTGGTTTCAG

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGATTCTGGGGTC

CCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATTACAGACGTGCAG

TGTGACGATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATGATGCCTTC

GGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCG

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MEXICAN INSTITUTE D £ THE PROPERTY

ccatctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgct ^ aat (SEQ ID NO: 61)

NO: 54:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGT

CAGGAGCAGCTGAAGGAGTCCGGAGGAGGCCTGGTAACGCCTGGAGGAACCCTGACACTCACC

TGCACAGCCTCTGGATTCTCCCTCAATGCCTACTACATGAACTGGGTCCGCCAGGCTCCAGGG

AAGGGGCTGGAATGGATCGGATTCATTACTCTGAATAATAATGTAGCTTACGCGAACTGGGCG

AAAGGCCGATTCACCTTCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACA

CCCGAGGACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTGCAATGGGTCGGTTG

GATCTCTGGGGCCATGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTC

TTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTC

AAGG (SEQ ID NO: 62).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 63; SEQ ID NO: 64 and SEQ ID NO: 65 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 53.

275

ρι £, 2. instituto μεχ · ολ :: ο

OF LA PRÜfiidAÍ)

INDUSTRIAL

In a further embodiment of the invention, the

polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 66; SEQ ID NO: 67 and SEQ ID NO:

that correspond to polynucleotides that encode the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 61 encoding the variable light chain region of SEQ ID NO: 53, the polynucleotide of SEQ ID NO: 62 encoding the variable heavy chain region of SEQ ID NO: 54; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65) of the variable light chain region of SEQ ID NO: 53; and the polynucleotides that encode the determining regions of

276

Complementarity (SEQ ID NO: 66; SEQ ID NO: 67 and SEQ ID NO: 68) of the variable heavy chain region of SEQ ID NO: 54.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCCCAAGTGCTGACCCAGACTCCATCGCCTGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAACTGCCAGGCCAGTCAGAGTGTTGATGATAACAACTGGTTAGGCTGGTATCAG

CAGAAACGAGGGCAGCCTCCCAAGTACCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTC

CCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAG

TGTGACGATGCTGCCACTTACTACTGTGCAGGCGGTTTTAGTGGTAATATCTTTGCTTTCGGC

GGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCA

TCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCT (SEQ

ID NO: 77)

NO: 70:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

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MEXICAN INDUSTRIAL EFFICIENCY INSTiT'JTU

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGTCTCTGGCTTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGAAAG

GGGCTGGAGTGGATCGGAATCATTGGTGGTTTTGGTACCACATACTACGCGACCTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAGAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAGGTGGTCCTGGTAATGGTGGTGACATCTGGGGC

CAAGGGACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCA

CCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACT (SEQ

ID NO: 78).

corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 69.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 82; SEQ ID NO: 83 and SEQ ID NO: 84 corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

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SEQ ID NO: 70.

The invention also includes polynucleotides that include one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 77 encoding the variable light chain region of SEQ ID NO: 69, the polynucleotide of SEQ ID NO: 78 encoding the variable heavy chain region of SEQ ID NO: 70; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 79; SEQ ID NO: 80 and SEQ ID NO: 81) of the region contemplated sequences of

<td>light chain</td><td>variable</td><td>of</td><td>the SEQ ID</td><td>NO: 69;</td><td>and</td><td>the</td>

<td>polynucleotides</td><td colspan="2">that encode</td><td>the regions</td><td colspan="2">determinants</td><td>of</td>

<td colspan="2">complementarity (SEQ ID</td><td>NOT:</td><td>82; SEQ ID NO:</td><td>83 and SEQ</td><td>ID</td><td>NOT:</td>

<td>84) from the region</td><td>chain</td><td colspan="2">variable weighing</td><td>the SEQ ID</td><td>NOT:</td><td> 70.</td>

The invention is further directed to polynucleotides encoding polypeptides of antibodies that possess IL-6 binding specificity. In one embodiment of the invention, the polynucleotides of the invention alternatively comprise or consist of the following sequence,

279

polynucleotides encoding the variable light chain polypeptide sequence of SEQ ID NO: 85:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGTACCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGTCCAGTCAGAGTGTTTATAATAATTTCTTATCGTGGTATCAGCAG

AAACCAGGGCAGCCTCCCAAGCTCCTGATCTACCAGGCATCCAAACTGGCATCTGGGGTCCCA

GATAGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGT

GACGATGCTGCCACTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCTTTCGGC

GGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCA

TCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTC (SEQ

ID NO: 93)

NO: 86:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACGCTCACCTGC

ACAGTCTCTGGAATCGACCTCAGTGACTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGAATCATTTATGCTGGTAGTGGTAGCACATGGTACGCGAGCTGGGCG

AAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACA

ACCGAGGACACGGCCACCTATTTCTGTGCCAGAGATGGATACGATGACTATGGTGATTTCGAT

CGATTGGATCTCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCA

TCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGC

CTGGTCAAGGACT (SEQ ID NO: 94).

280

In a further embodiment of the '' ^ iWfencTónT ^ the polynucleotides encoding fragments of the antterrerpΰ possessing specificity for binding to IL-6 comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 95; SEQ ID NO: 96 and SEQ ID NO:

corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 85.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 98; SEQ ID NO: 99 and SEQ ID NO: 100 which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td>heavy chain</td><td>variable of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

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INSTITUTO MEXICANO DE LA PAODEDA INDUSTRIAL the following polynucleotides encoding antibody fragments: the polynucleotide of SEQ ID NO: 93 encoding the variable light chain region of SEQ ID NO: 85, the polynucleotide of SEQ ID NO: 94 which encodes the variable heavy chain region of SEQ ID NO: 86; polynucleotides encoding complementarity determining regions (SEQ ID NO: 95; SEQ ID NO: 96 and SEQ ID NO: 97) of the variable light chain region of SEQ ID NO: 85; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 98; SEQ ID NO: 99 and SEQ ID NO: 100) of the variable heavy chain region of SEQ ID NO:

86.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAAATGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAA

TCAGGGCAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCG

CGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCC

GATGCTGCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCTTTCGGC

282

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GGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCA

TCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTC (SEQ

ID NO: 109)

NO: 102:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCTCAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACAGCCTCTGGATTCTCCCTCAGTAACTACTACATGACCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGAATGATTTATGGTAGTGATGAAACAGCCTACGCGAACTGGGCGATA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCC

GCGGACACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAAC

TTGTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTC

CCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAG

G (SEQ ID NO: 110).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 111; SEQ ID NO: 112 and SEQ ID NO: 113 corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

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SEQ ID NO: 101.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 114; SEQ ID NO: 115 and SEQ ID NO: 116 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td colspan="4">of the variable heavy chain sequence of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 109 encoding the variable light chain region of SEQ ID NO: 101, the polynucleotide of SEQ ID NO: 110 encoding the variable heavy chain region of SEQ ID NO: 102; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 111; SEQ ID NO: 112 and SEQ ID NO: 113) of the variable light chain region of SEQ ID NO:

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101; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 114; SEQ ID NO:

115 and SEQ ID NO: 116) of the variable heavy chain region of SEQ ID NO: 102.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCAGCCGTGCTGACCCAGACACCATCACOCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAGTTGCCAGTCCAGTCAGAGTGTTGGTAATAACCAGGACTTATCCTGGTTTCAG

CAGAGACCAGGGCAGCCTCCCAAGCTCCTGATCTACGAAATATCCAAACTGGAATCTGGGGTC

CCATCGCGGTTCAGCGGCAGTGGATCTGGGACACACTTCACTCTCACCATCAGCGGCGTACAG

TGTGACGATGCTGCCACTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCT (SEQ ID NO: 130)

NO: 123

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CACTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

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ACAGTCTCTGGATTCTCCCTCAGTAGTCGTACAATGTCCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAGTGGATCGGATACATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGATTGGGCGATACTGGTGGTCACGCTTATGCTACT

CGCTTAAATCTC (SEQ ID NO: 131).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 132; SEQ ID NO: 133 and SEQ ID NO: 134 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 122.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 135; SEQ ID NO: 136 and SEQ ID NO: 137 which correspond to polynucleotides that encode the complementarity determining regions (CDRs, or regions

<td colspan="2">hypervariables) of the sequence</td><td colspan="2">variable heavy chain</td><td>the</td>

<td>The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides that</td><td>include</td><td>one or more</td><td>sequences</td><td>of</td>

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polynucleotides encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of antibody: the polynucleotide of SEQ ID NO: 130 encoding the variable light chain region of SEQ ID NO: 122, the polynucleotide of SEQ ID NO: 131 encoding the variable heavy chain region of SEQ ID NO: 123; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 132; SEQ ID NO: 133 and SEQ ID NO: 134) of the variable light chain region of SEQ ID NO: 122; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 135; SEQ ID NO: 136 and SEQ ID NO: 137) of the variable heavy chain region of SEQ ID NO: 123.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

287

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INSTITUTO MEXICANO yGCCACATTTGCAGCCGTGCTGACCCAGACACCATCGTCCGTGTCTG ^ eTGTSSSftGGCACA gtcagcatcagttgccagtccagtcagagtgtttatagtaataastacctagcc: tggtatc.ag

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACTGGACATCCAAACTGGCATCTGGGGCC

CCATCACGGTTCAGCGGCAGTGGATCTGGGACACAATTCACTCTCACCATCAGCGGCGTGCAG

TGTGACGATGCTGCCACTTACTACTGTCTAGGCGCTTATGATGATGATGCTGATAATGCT (SEQ ID NO: 146)

NO: 139:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAAGAGTCCGGGGGTCGCCTGGTCAAGCCTGACGAAACCCTGACACTCACCTGC

ACAGCCTCTGGATTCTCCCTGGAGGGCGGCTACATGACCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGAATCAGTTATGATAGTGGTAGCACATACTACGCGAGCTGGGCGAAA

GGCCGATTCACCATCTCCAAGACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACA

ACCGAGGACACGGCCACCTATTTCTGCGTCAGATCACTAAAATATCCTACTGTTACTTCTGAT

GACTTG (SEQ ID NO: 147).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 148; SEQ ID NO: 149 and SEQ ID NO: 150 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

288

MEXICAN INSTITUTE OF THE? PO? 1EDAD (hypervariables) of the string sequence ligetfSforWfiát

SEQ ID NO: 138. -

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more poiinucleotide sequences of SEQ ID NO 151; SEQ ID NO: 152 and SEQ ID NO: 153 corresponding to poiinucleotides that encode complementarity determining regions (CDRs, or regions

<td colspan="2">hypervariables) of the</td><td>sequence of</td><td colspan="3">variable heavy chain the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>poiinucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>poiinucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, poiinucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following poiinucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 146 encoding the variable light chain region of SEQ ID NO: 138, the polynucleotide of SEQ ID NO: 147 encoding the variable heavy chain region of SEQ ID NO: 139; the polynucleotides that encode the complementarity determining regions (SEQ ID NO: 148; SEQ ID NO: 149 and SEQ ID NO:

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150) of the variable light chain regionINI knowYou^ a NO:

138; and the polynucleotides that cocTltióSli “Tas ..... complementary determining regimes (SEQ ID NO: 151; SEQ ID NO:

152 and SEQ ID NO: 153) of the variable heavy chain region of SEQ ID NO: 139.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCAGCCGTGCTGACCCAGACACCATCACCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAGTTGCCAGTCCAGTCAGAGTGTTTATAATAATAACGACTTAGCCTGGTATCAG

CAGAAACCAGGGCAGCCTCCTAAACTCCTGATCTATTATGCATCCACTCTGGCATCTGGGGTC

CCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAG

TGTGACGATGCTGCCGCTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCT (SEQ ID NO: 162)

NO: 155:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

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CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGTATCTGGATTATCCCTCAGTAGCAATACAATAAACTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAGTGGATCGGATACATTTGGAGTGGTGGTAGTACATACTACGCGAGCTGGGTGAAT

GGTCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAGGGGGTTACGCTAGTGGTGGTTATCCTTATGCC

ACTCGGTTGGATCTC (SEQ ID NO: 163).

NO: 166 which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 154.

NO: 169 corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 155.

The invention also contemplates sequences of

291

WRONG polynucleotides that include one more

INDUSTRIAL polynucleotide sequences encoding antibody fragments described herein. In an embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 162 encoding the variable light chain region of SEQ ID NO: 154, the polynucleotide of SEQ ID NO: 163 encoding the variable heavy chain region of SEQ ID NO: 155; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 164; SEQ ID NO: 165 and SEQ ID NO: 166) of the variable light chain region of SEQ ID NO: 154; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 167; SEQ ID NO: 168 and SEQ ID NO: 169) of the variable heavy chain region of SEQ ID NO: 155.

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ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCAGCCGTGCTGACCCAGACACCATCCTCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAATTGCCAGTCCAGTCAGAGTGTTTATAATAACGACTACTTATCCTGGTATCAA

CAGAGGCCAGGGCAACGTCCCAAGCTCCTAATCTATGGTGCTTCCAAACTGGCATCTGGGGTC

CCGTCACGGTTCAAAGGCAGTGGATCTGGGAAACAGTTTACTCTCACCATCAGCGGCGTGCAG

TGTGACGATGCTGCCACTTACTACTGTCTGGGCGATTATGATGATGATGCTGATAATACT (SEQ ID NO: 178)

NO: 171:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGACACTCACTTGC

ACAGTCTCTGGATTCACCCTCAGTACCAACTACTACCTGAGCTGGGTCCGCCAGGCTCCAGGG

AAGGGGCTAGAATGGATCGGAATCATTTATCCTAGTGGTAACACATATTGCGCGAAGTGGGCG

AAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCCG

ACAACCGAGGACACAGCCACGTATTTCTGTGCCAGAAATTATGGTGGTGATGAAAGTTTG (SEQ ID NO: 179).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 180; SEQ ID NO: 181 and SEQ ID NO: 182 corresponding to polynucleotides encoding the

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MEXICAN INSTITUTE V'ítíG- '-'C'í M DE LA PROPicDAO VV-ütL f j- /

INDUSTRIAL Complementarity-determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of

SEQ ID NO: 170.

In a further embodiment of the invention, the polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more poiinucleotide sequences of SEQ ID NO 183; SEQ ID NO: 184 and SEQ ID

NO: 185 corresponding to poiinucleotides that encode the complementary determining regions (CDR, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td>heavy chain</td><td>variable of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>poiinucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>poiinucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, poiinucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following poiinucleotides encoding fragments of antibody: the polynucleotide of SEQ ID NO: 178 encoding the variable light chain region of SEQ ID NO: 170, the polynucleotide of SEQ ID NO: 179 encoding the variable heavy chain region of SEQ ID NO: 171; the polynucleotides that encode the determining regions of

294

Complementarity (SEQ ID NO: 180; SEQ ID NO: 181 and SEQ ID NO:

182) of the variable light chain region of SEQ ID NO:

170; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 183; SEQ ID NO:

184 and SEQ ID NO: 185) of the variable heavy chain region of SEQ ID NO: 171.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGAGCCA

GTCACCATCAAGTGCCAGGCCAGTGAGACCATTGGCAATGCATTAGCCTGGTATCAGCAGAAA

TCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCAAACTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGCC

GATGCTGCCACTTACTACTGTCAATGGTGTTATTTTGGTGATAGTGTT (SEQ ID NO:

194)

NO: 187:

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ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCACTGTGCTCA ^ AGGTGTCCÁGTGT

CAGGAGCAGCTGGTGGAGTCCGGGGGAGGCCTGGTCCAGCCTGAGGGATCCCTGACACTCACCC

TGCACAGCCTCTGGATTCGACTTCAGTAGCGGCTACTACATGTGCTGGGTCCGCCAGGCTCCA

GGGAAGGGGCTGGAGTGGATCGCGTGTATTTTCACTATTACTACTAACACTTACTACGCGAGC

TGGGCGAAAGGCCGATTCACCATCTCCAAGACCTCGTCGACCACGGTGACTCTGCAAATGACC

AGTCTGACAGCCGCGGACACGGCCACCTATCTCTGTGCGAGAGGGATTTATTCTGATAATAAT

TATTATGCCTTG (SEQ ID NO: 195).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 196; SEQ ID NO: 197 and SEQ ID NO: 198 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 186.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 199; SEQ ID NO: 200 and SEQ ID NO: 201 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 187.

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The invention also

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contemplates polynucleotide sequences including one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucieotide of SEQ ID NO: 194 encoding the variable light chain region of SEQ ID NO: 186, the polynucieotide of SEQ ID NO: 195 encoding the variable heavy chain region of SEQ ID NO: 187; polynucleotides that encode complementarity determining regions (SEQ ID NO: 196; SEQ ID NO: 197 and SEQ ID NO:

198) of the variable light chain region of SEQ ID NO:

186; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 199; SEQ ID NO: 200 and SEQ ID NO: 201) of the variable heavy chain region of SEQ ID NO: 187.

The invention is further directed to polynucleotides encoding polypeptides of antibodies that possess IL-6 binding specificity. In one embodiment of the invention, the poiinucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the polypeptide sequence of

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INDUSTRIAL variable light chain of SEQ ID NO: 202:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGAGCCA

GTCACCATCAAGTGCCAGGCCAGTGAGAGCATTGGCAATGCATTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGCC

GATGCTGCCGCTTACTACTGTCAATGGTGTTATTTTGGTGATAGTGTT (SEQ ID NO:

210) In another embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID

NO: 203:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGCAGCAGCTGGTGGAGTCCGGGGGAGGCCTGGTCAAGCCGGGGGCATCCCTGACACTCACCC

TGCAAAGCCTCTGGATTCTCCTTCAGTAGCGGCTACTACATGTGCTGGGTCCGCCAGGCTCCA

GGGAAGGGGCTGGAGTCGATCGCATGCATTTTTACTATTACTGATAACACTTACTACGCGAAC

TGGGCGAAAGGCCGATTCACCATCTCCAAGCCCTCGTCGCCCACGGTGACTCTGCAAATGACC

AGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGGGGGATTTATTCTACTGATAAT

TATTATGCCTTG (SEQ ID NO: 211).

298

NO: 214 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 202.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 215; SEQ ID NO: 216 and SEQ ID NO: 217 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td>heavy chain</td><td>variable of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

299

polynucleotides encoding the complementarity determining regions (SEQ ID NO: 212; SEQ ID NO: 213 and SEQ ID NO: 214) of the variable light chain region of SEQ ID NO: 202; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 215; SEQ ID NO: 216 and SEQ ID NO: 217) of the variable heavy chain region of SEQ ID NO: 203.

The invention is further directed to polynucleotides encoding polypeptides of antibodies that possess IL-β binding specificity. In an embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the variable light chain polypeptide sequence of SEQ ID NO: 218:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGAGCCA

GTCACCATCAAGTGCCAGGCCAGTCAGAGCGTTAGTAGCTACTTAAACTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTACAGGGCATCCACTCTGGAATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCC

GATGCTGCCACTTACTACTGTCAATGTACTTATGGTACTAGTAGTAGTTATGGTGCTGCT (SEQ ID NO: 226)

300

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

NO: 219:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACCGTCTCTGGTATCTCCCTCAGTAGCAATGCAATAAGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGAATCATTAGTTATAGTGGTACCACATACTACGCGAGCTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATCACTAGTCCGACA

ACCGAGGACACGGCCACCTACTTCTGTGCCAGAGATGACCCTACGACAGTTATGGTTATGTTG

ATACCTTTTGGAGCCGGCATGGACCTC (SEQ ID NO: 227).

In a further embodiment of the invention, the 10 polynucleotides encoding fragments of the antibody possessing IL-β binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 228; SEQ ID NO: 229 and SEQ ID NO: 230 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 218.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 231; SEQ ID NO: 232 and SEQ ID NO: 233 corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

301

IΜ ΡI 63 ^ *

MEXICAN INSTITUTE

PROPERTY VV- ^ CSjL1^”

INDUSTRIAL

SEQ ID NO: 219.

The invention contemplates also polynucleotide sequences that include one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 226 encoding the variable light chain region of SEQ ID NO: 218, the polynucleotide of SEQ ID NO: 227 encoding the variable heavy chain region of SEQ ID NO: 219; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 228; SEQ ID NO: 229 and SEQ ID NO: 230) of the variable light chain region of SEQ ID NO:

218; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 231; SEQ ID NO: 232 and SEQ ID NO: 233) of the variable heavy chain region of SEQ ID NO: 219.

302

EMPI Mexican Institute of INDUSTRIAL PROPERTY

polynucleotides encoding the variable light chain polypeptide sequence of SEQ ID NO: 234:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCCCAAGTGCTGACCCAGACTGCATCGCCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAACTGCCAGGCCAGTCAGAGTGTTTATAAGAACAACTACTTATCCTGGTATCAG

CAGAAACCAGGGCAGCCTCCCAAAGGCCTGATCTATTCTGCATCGACTCTAGATTCTGGGGTC

CCATTGCGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAG

TGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTAT

GCT (SEQ ID NO: 242)

NO: 235:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGAGGGATCCCTGACACTCACCTGC

ACAGCCTCTGGATTCTCCTTCAGTAGCTACTGGATGTGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAGTGGATCGCATGCATTGTTACTGGTAATGGTAACACTTACTACGCGAACTGGGCG

AAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTG

ACAGCCGCGGACACGGCCACCTATTTTTGTGCGAAAGCCTATGACTTG (SEQ ID NO:

243) .

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more sequences of

303

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY polynucleotides of SEQ ID NO 244; SEQ ID NO: 245 and SEQ ID NO: 246 corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 234.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 247; SEQ ID NO: 248 and SEQ ID NO: 249 which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td>heavy chain</td><td>variable of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

304 Nu / L3.

SEQ ID NO variable heavy chain:

IMPI

ΙΝΓΓΠΊΙΤΟ MFX.T.'Nf)

MEXICAN INSTITUTE OF L ?. ? 3G?! INDOS'XÍ / .L AGE

235; the polynucleotides that encode the complementarity determining regions (SEQ ID NO: 244; SEQ ID NO: 245 and SEQ ID NO:

246) of the variable light chain region of SEQ ID NO: 234; and the poiinucleotides encoding the complementarity determining regions (SEQ ID NO: 247; SEQ ID NO: 248 and SEQ ID NO: 249) of the variable heavy chain region of SEQ ID NO: 235.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

TCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACA

GTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATGACAACAACTATTTATCCTGGTATCAG

CAGAAACCAGGACAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTC

CCATCGCGGTTCAAAGGCACGGGATCTGGGACACAGTTCACTCTCACCATCACAGACGTGCAG

TGTGACGATGCTGCCACTTACTATTGTGCAGGCGTTTTTAATGATGATAGTGATGATGCC (SEQ ID NO: 258)

In another embodiment of the invention, the poiinucleotides of the invention comprise or, alternatively, consist of the following sequence of poiinucleotides encoding the

305

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IbJ7I rT? Variable variable heavy chain polypeptide sequence of SEQ ID

NO: 251:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCCCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACACTCTCTGGATTCTCCCTCAGTGCATACTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGATTCATTACTCTGAGTGATCATATATCTTACGCGAGGTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTGCAATGGGTCGGTTGGAT

CTC (SEQ ID NO: 259).

NO: 262 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 250.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 263; SEQ ID NO: 264 and SEQ ID NO: 265 which correspond to polynucleotides that encode the complementarity determining regions (CDRs, or regions

306

JE * INDUSTRIAL PROPERTY hypervariables) of the variable heavy chain sequence of the

SEQ ID NO: 251

The invention also contemplates polynucleotides that include one or more polynucleotides that encode fragments sequence of antibody sequences described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 258 encoding the variable light chain region of SEQ ID NO: 250, the polynucleotide of SEQ ID NO: 259 encoding the variable heavy chain region of SEQ ID NO: 251; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 260; SEQ ID NO: 261 and SEQ ID NO: 262) of the variable light chain region of SEQ ID NO: 250; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 263; SEQ ID NO: 264 and SEQ ID NO: 265) of the variable heavy chain region of SEQ ID NO: 251.

307

INSTITUTO MEXICANO alternative way, consist of the following ^^ or ^ Úen ^ ra ^ ae polynucleotides that encode the sequence Πδ poTipept'lClOS CtS- variable light chain of SEQ ID NO: 266:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTCGCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGCGGCTGTGGGAGGCACA

GTCACCArCAGTTGCCAGGCCAGTCAGAGTGTTTATAACAACAAAAATTTAGCCTGGTATCAG

CAGAAATCAGGGCAGCCTCCCAAGCTCCTGATCTACTGGGCATCCACTCTGGCATCTGGGGTC

TCATCGCGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCGTCAGCGGCGTGCAG

TGTGACGATGCTGCCACTTACTACTGTCTAGGCGTTTTTGATGATGATGCTGATAATGCT (SEQ ID NO: 274)

NO: 267:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAATGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGCCTCTGGATTCTCCCTCAGTAGCTACTCCATGACCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATATATCGGAGTCATTGGTACTAGTGGTAGCACATACTACGCGACCTGGGCGAAA

GGCCGATTCACCATCTCCAGAACCTCGACCACGGTGGCTCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGTCAGGAGTCTTTCTTCTATTACTTTCTTG (SEQ ID

NO: 275).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or are

308

IMPIOS

INSTITUTO MEXICANO alternative, consist of one or more polynucleotide sequences of SEQ ID NO 27 6; SEQ ID N'O: 'Σ7Τ ~' ΉΈ ^ ΊΤΓ ”NO: 278 which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 266.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 279; SEQ ID NO: 280 and SEQ ID NO: 281 which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

309

polynucieotide of SEQ ID NO: 275 encoding ^ iia'iuiie variable heavy chain of SEQ IB - NO-i-? 6 '7; 1 = polynucleotides that encode the complementarity determining regions (SEQ ID NO: 276; SEQ ID NO: 277 and SEQ ID NO:

278) of the variable light chain region of SEQ ID NO:

266; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 279; SEQ ID NO:

280 and SEQ ID NO: 281) of the variable heavy chain region of SEQ ID NO: 267.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCATTCGAATTGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACA

GTCACCATCAATTGCCAGGCCAGTCAGAACATTTATAGATACTTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGTTCCTGATCTATCTGGCATCTACTCTGGCATCTGGGGTCCCATCG

CGGTTTAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCC

GATGCTGCCACTTACTACTGTCAAAGTTATTATAGTAGTAATAGTGTCGCT (SEQ ID NO:

290)

In another embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of

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the following polynucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID

NO: 283:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGGAGCAGCTGGTGGAGTCCGGGGGAGACOTGGTCCAGCCTGAGGGATCCCTGACACTCACC

TGCACAGCTTCTGAGTTAGACTTCAGTAGCGGCTACTGGATATGCTGGGTCCGCCAGGTTCCA

GGGAAGGGGCTGGAGTGGATCGGATGCATTTATACTGGTAGTAGTGGTAGCACTTTTTACGCG

AGTTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATG

ACCAGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGAGGTTATAGTGGCTTTGGT

TACTTTAAGTTG (SEQ ID NO: 291).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 292; SEQ ID NO: 293 and SEQ ID NO: 294 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 282.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 295; SEQ ID NO: 296 and SEQ ID NO: 297 corresponding to polynucleotides encoding the

311

MEXICAN INSTITUTE

DELA PROPIEDAD VV --- tíSLlS / industrial v<ú2AC2 £ * '' Complementarity-determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

<td>the light chain region</td><td>variable</td><td>of</td><td>SEQ ID NO: 282,</td><td>the</td>

<td>polynucleotide of SEQ ID</td><td>NO: 291</td><td>than</td><td>encode the region</td><td>of</td>

<td>variable heavy chain</td><td>of the</td><td>I KNOW THAT</td><td>ID NO: 283;</td><td>the</td>

<td colspan="2">polynucleotides that encode</td><td colspan="2">determining regions</td><td>of</td>

Complementarity (SEQ ID NO: 292; SEQ ID NO: 293 and SEQ ID NO:

294) of the variable light chain region of SEQ ID NO:

282; and the polynucleotides encoding the complementarity determining regions (SEQ ID NO: 295; SEQ ID NO: 296 and SEQ ID NO: 297) of the variable heavy chain region of SEQ ID NO: 283.

The invention is further directed to polynucleotides encoding polypeptides of antibodies that possess IL-6 binding specificity. In one form of the

312

Invention, the polynucleotides of the invention comprise or alternatively consist of the following polynucleotide sequence encoding the variable light chain polypeptide sequence of SEQ ID NO: 298:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGGCCAGTGAGGACATTTATAGGTTATTGGCCTGGTATCAACAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGATTCATCCGATCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCGCCATCAGCGGTGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTCAACAGGCTTGGAGTTATAGTGATATTGATAATGCT (SEQ

ID NO: 306)

NO: 299:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCGGGGACACCCCTGACACTCACCTGC

ACAGCCTCTGGATTCTCCCTCAGTAGCTACTACATGAGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGAATCATTACTACTAGTGGTAATACATTTTACGCGAGCTGGGCGAAA

GGCCGGCTCACCATCTCCAGAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAACTTCTGATATTTTTTATTATCGTAACTTG (SEQ ID NO: 307).

In a further embodiment of the invention, the polynucleotides encoding fragments of the antibody that

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INDUSTRIAL -j:

possesses IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 308; SEQ ID NO: 309 and SEQ ID NO: 310 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 298.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 311; SEQ ID NO: 312 and SEQ ID NO: 313 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

314

<td>the light chain region</td><td>variable</td><td>of</td><td colspan="2">SEQ ID NO: 298,</td><td>the</td>

<td>polynucleotide of SEQ ID</td><td>NO: 307</td><td>than</td><td>encode</td><td>the region</td><td>of</td>

<td>variable heavy chain</td><td>of the</td><td>I KNOW THAT</td><td>ID NO:</td><td> 2 99;</td><td>the</td>

<td colspan="2">polynucleotides that encode</td><td colspan="3">determining regions</td><td>of</td>

Complementarity (SEQ ID NO: 308; SEQ ID NO: 309 and SEQ ID NO:

310) of the variable light chain region of SEQ ID NO:

298; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 311; SEQ ID NO: 312 and SEQ ID NO: 313) of the variable heavy chain region of SEQ ID NO: 299.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACGTTTGCAGCCGTGCTGACCCAGACTGCATCACCCGTGTCTGCCGCTGTGGGAGCCACA

GTCACCATCAACTGCCAGTCCAGTCAGAGTGTTTATAATGACATGGACTTAGCCTGGTTTCAG

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTC

CCATCGCGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGCAG

TGTGACGATGCTGCCACTTACTACTGTCTAGGCGCTTTTGATGATGATGCTGATAATACT (SEQ ID NO: 322)

In another embodiment of the invention, the polynucleotides of

315

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

_ , , z , IPVUJ I iu / ll. > 1 - the invention comprises or alternatively consists of the following polynucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID

NO: 315

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGTCTCTGGATTCTCCCTCACTAGGCATGCAATAACCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGATGCATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTCAGAATCACCAGTCCGACAACC

GAGGACACGGCCACCTACTTCTGTGCCAGAGTCATTGGCGATACTGCTGGTTATGCTTATTTT

ACGGGGCTTGACTTG (SEQ ID NO: 323).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 324; SEQ ID NO: 325 and SEQ ID NO: 326 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 314.

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

NO: 329 that correspond to polynucleotides that encode the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td colspan="3">variable heavy chain sequence of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one. or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 322 encoding the variable light chain region of SEQ ID NO: 314, the polynucleotide of SEQ ID NO: 323 encoding the variable heavy chain region of SEQ ID NO: 315; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 324; SEQ ID NO: 325 and SEQ ID NO: 326) of the variable light chain region of SEQ ID NO: 314; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 327; SEQ ID NO: 328 and SEQ ID NO: 329) of the variable heavy chain region of SEQ ID NO: 315.

The invention is further directed to polynucleotides encoding polypeptides of antibodies that possess a

317

IMPIOUS

MEXICAN INSTITUTE V OF PROPERTY V INDUSTRIAL specificity of binding to IL-6. In an embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the variable light chain polypeptide sequence of SEQ ID NO: 330:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGGCCAGTCAGAGTGTTTATAATTGGTTATCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATACTGCATCCAGTCTGGCATCTGGGGTCCCATCG

CGGTTCAGTGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGGAGTGTGCC

GATGCTGCCACTTACTACTGTCAACAGGGTTATACTAGTGATGTTGATAATGTT (SEQ ID

NO: 338)

NO: 331:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGGCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGACACTCACCTGC

ACAGTCTCTGGAATCGACCTCAGTAGCTATGCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATACATCGGAATCATTAGTAGTAGTGGTAGCACATACTACGCGACCTGGGCGAAA

GGCCGATTCACCATCTCACAAGCCTCGTCGACCACGGTGGATCTGAAAATTACCAGTCCGACA

ACCGAGGACTCGGCCACATATTTCTGTGCCAGAGGGGGTGCTGGTAGTGGTGGTGTTTGGCTG

CTTGATGGTTTTGATCCC (SEQ ID NO: 339).

In a further embodiment of the invention, the

318

NO: 342 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 330.

In a further embodiment of the invention, the 10 polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 343; SEQ ID NO: 344 and SEQ ID NO: 345 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td>heavy chain</td><td>variable of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

319

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MEXICAN INSTITUTE PE THE PROPERTY

INDUSTRIAL antibody: the polynucleotide of SEQ ID NO: 338 encoding the variable light chain region of SEQ ID NO: 330, the polynucleotide of SEQ ID NO: 339 encoding the variable heavy chain region of SEQ ID NO : 331; polynucleotides that encode complementarity determining regions (SEQ ID NO: 340; SEQ ID NO: 341 and SEQ ID NO:

342) of the variable light chain region of SEQ ID NO:

330; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 343; SEQ ID NO:

344 and SEQ ID NO: 345) of the variable heavy chain region of SEQ ID NO: 331.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAAATGTGCCGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGC

ACAGTCACCATCAATTGCCAGGCCAGTGAGAACATTTATAATTGGTTAGCCTGGTATCAGCAG

AAACCAGGGCAGCCTCCCAAGCTCCTGATCTATACTGTAGGCGATCTGGCATCTGGGGTCTCA

TCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGT

GCCGATGCTGCCACTTACTATTGTCAACAGGGTTATAGTAGTAGTTATGTTGATAATGTT (SEQ ID NO: 354)

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In another embodiment of the iPiUUSTiUAL invention the polynucleotides of the invention comprise or, alternatively, consist of the following poiinucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID

NO: 347:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACC

TGCACAGTCTCTGGATTCTCCCTCAATGACTATGCAGTGGGCTGGTTCCGCCAGGCTCCAGGG

AAGGGGCTGGAATGGATCGGATACATTCGTAGTAGTGGTACCACAGCCTACGCGACCTGGGCG

AAAGGCCGATTCACCATCTCCGCTACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACA

ACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGGTGCTGGTAGTAGTGGTGTGTGGATC

CTTGATGGTTTTGCTCCC (SEQ ID NO: 355).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more poiinucleotide sequences of SEQ ID NO 356; SEQ ID NO: 357 and SEQ ID NO: 358 which correspond to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 346.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more sequences of

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INDUSTRIAL _ polynucleotides of SEQ ID NO 359; SEQ ID NO: 360 and SEQ ID

NO: 361 corresponding to polynucleotides that encode the complementary determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 347.

The invention of sequences also contemplates polynucleotides that include one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 354 encoding the variable light chain region of SEQ ID NO: 346, the polynucleotide of SEQ ID NO: 355 encoding the variable heavy chain region of SEQ ID NO: 347; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 356; SEQ ID NO: 357 and SEQ ID NO: 358) of the variable light chain region of SEQ ID NO: 346; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 359; SEQ ID NO: 360 and SEQ ID NO: 361) of the variable heavy chain region of SEQ ID NO: 347.

The invention is further directed to polynucleotides

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INDUSTRIAL encoding polypeptides of antibodies that possess an, i | IL-6 binding specificity. In one embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the variable light chain polypeptide sequence of SEQ ID NO: 362:

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCTCAAGTGCTGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCAGAACAACTACTTATCCTGGTTTCAG

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCGGCCACTCTGGCATCTGGGGTC

CCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTOACCATCAGCGACCTGGAG

TGTGACGATGCTGCCACTTACTACTGTGCAGGCGCTTATAGGGATGTGGATTCT (SEQ ID

NO: 370)

NO: 363:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGGGGCATCCCTGACACTCACCTGC

ACAGCCTCTGGATTCTCCTTTACTAGTACCTACTACATCTACTGGGTCCGCCAGGCTCCAGGG

AAGGGGCTGGAGTGGATCGCATGTATTGATGCTGGTAGTAGTGGTAGCACTTACTACGCGACC

TGGGTGAATGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACC

AGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAAATGGGATTATGGTGGTAATGTT

GGTTGGGGTTATGACTTG (SEQ ID NO: 371).

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NO: 374 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 362.

NO: 377 that correspond to polynucleotides that encode the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td colspan="2">of the sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In an embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all

324

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MEXICAN INSTITUTE OF PROPERTY. 'L · INDUSTRIAL

the following polynucleotides that encode antibody fragments: the polynucleotide of SEQ ID NO: 370 that encodes

<td colspan="2">the light chain region</td><td colspan="3">SEQ ID variable</td><td>NO: 362,</td><td>the</td>

<td>polynucleotide</td><td>of the SEQ ID</td><td>NO: 371</td><td>than</td><td>encode</td><td>the region</td><td>of</td>

<td>heavy chain</td><td>variable</td><td>of the</td><td>I KNOW THAT</td><td>ID NO:</td><td> 363;</td><td>the</td>

<td>polynucleotides</td><td colspan="2">that encode the</td><td colspan="3">determining regions</td><td>of</td>

Complementarity (SEQ ID NO: 372; SEQ ID NO: 373 and SEQ ID NO: 374) of the variable light chain region of SEQ ID NO: 362; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 375; SEQ ID NO: 376 and SEQ ID NO: 377) of the variable heavy chain region of SEQ ID NO: 363.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCATTCGAATTGACCCAGACTCCATCCTCCGTGGAGGCAGCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGGCCAGTCAGAGCATTAGTAGTTACTTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGTTCCTGATCTACAGGGCGTCCACTCTGGCATCTGGGGTCCCATCG

CGATTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCC

GATGCTGCCACTTACTACTGTCAAAGCTATTATGATAGTGTTTCAAATCCT (SEQ ID NO:

325

386)

NO: 379:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGAGGGATCCCTGACACTCACCTGC

AAAGCCTCTGGACTCGACCTCGGTACCTACTGGTTCATGTGCTGGGTCCGCCAGGCTCCAGGG

AAGGGGCTGGAGTGGATCGCTTGTATTTATACTGGTAGTAGTGGTTCCACTTTCTACGCGAGC

TGGGTGAATGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACC

AGTCTGACAGCCGCGGACACGGCCACTTATTTTTGTGCGAGAGGTTATAGTGGTTATGGTTAT

TTTAAGTTG (SEQ ID NO: 387).

NO: 390 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 378.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or are

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INSTITUTO MEX! ~ V: .3 OF INDUSTRIAL PROPERTY

alternatively, they consist of one or more polynucleotide sequences of SEQ ID NO 391; SEQ ID NO: 392 and SEQ ID

NO: 393 that correspond to polynucleotides that encode the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td colspan="2">of the sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">by antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three, or more, including all of the following polynucleotides encoding fragments of antibody: the polynucleotide of SEQ ID NO: 386 encoding the variable light chain region of SEQ ID NO: 378, the polynucleotide of SEQ ID NO: 387 encoding the variable heavy chain region of SEQ ID NO: 379; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 388; SEQ ID NO: 389 and SEQ ID NO: 390) of the variable light chain region of SEQ ID NO: 378; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 391; SEQ ID NO:

392 and SEQ ID NO: 393) of the variable heavy chain region of SEQ ID NO: 379.

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INDUSTRIAL -1

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GTCACATTTGCCATCGAAATGACCCAGAGTCCATTCTCCGTGTCTGCAGCTGTGGGAGGCACA

GTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATAAGAACAACCAATTATCCTGGTATCAG

CAGAAATCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCGGCTCTGGCATCTGGGGTC

CCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACGTGCAG

TGTGACGATGCTGCCACTTACTACTGTGCAGGCGCTATTACTGGTAGTATTGATACGGATGGT (SEQ ID NO: 402)

NO: 395:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGGGGCATCCCTGACACTCACCTGC

ACAACTTCTGGATTCTCCTTCAGTAGCAGCTACTTCATTTGCTGGGTCCGCCAGGCTCCAGGG

AAGGGGCTGGAGTGGATCGCATGCATTTATGGTGGTGATGGCAGCACATACTACGCGAGCTGG

GCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACGCTGCAAATGACCAGT

CTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGAGAATGGGCATATAGTCAAGGTTAT

328

TTTGGTGCTTTTGATCTC (SEQ ID NO: 403).

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 404; SEQ ID NO: 405 and SEQ ID NO: 406 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 394.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 407; SEQ ID NO: 408 and SEQ ID NO: 409 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td colspan="2">heavy chain sequence</td><td colspan="2">variable of the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or in a manner

329

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INST1TUTJ Μ £?;: - 2ΛΝΟ D & LA? XD? ÍEDAD INDUSTRIAL

Alternative consist of one, two, three or more, including all of the following polynucleotides encoding antibody fragments: the polynucleotide of SEQ ID NO: 402 encoding the variable light chain region of SEQ ID NO: 394, the polynucleotide from SEQ ID NO: 403 encoding the variable heavy chain region of SEQ ID NO: 395; polynucleotides that encode complementarity determining regions (SEQ ID NO: 404; SEQ ID NO: 405 and SEQ ID NO: 406) of the variable light chain region of SEQ ID NO: 394; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 407; SEQ ID NO: 408 and SEQ ID NO: 409) of the variable heavy chain region of SEQ ID NO: 395.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGAGCCA

GTCACCATCAAGTGCCAGGCCAGTGAGGATATTAGTAGCTACTTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGCTGCATCCAATCTGGAATCTGGGGTCTCATCG

CGATTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGCC

330

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MEXICAN INSTITUTE CE LA PÉ.Í'I'IlDAD

INDUSTRIAL

GATGCTGCCACCTATTACTGTCAATGTACTTATGGTACTATTTCTATTAGTGATGGTAATGCT (SEQ ID NO: 418)

NO: 411:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAATGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGTCTCTGGATTCTCCCTCAGTAGCTACTTCATGACCTGGGTCCGCCAGGCTCCAGGGGAG

GGGCTGGAATACATCGGATTCATTAATCCTGGTGGTAGCGCTTACTACGCGAGCTGGGTGAAA

GGCCGATTCACCATCTCCAAGTCCTCGACCACGGTAGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGGGTTCTGATTGTTTCTTATGGAGCCTTTACCATC (SEQ ID NO: 419).

NO: 422 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 410.

In a further embodiment of the invention, the polynucleotides encoding fragments of the antibody that

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MEXICAN INSTITUTE tt-· ,·</

DE la p :: 3.'U). · .-> \

INDUSTIUAL '------ possesses an IL-6 binding specificity comprising, or alternatively, consisting of one or more poiinucleotide sequences of SEQ ID NO 423; SEQ ID NO: 424 and SEQ ID

NO: 425 that correspond to poiinucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 411.

The invention also contemplates poiinucleotide sequences that include one or more poiinucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, poiinucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following poiinucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 418 encoding the variable light chain region of SEQ ID NO: 410, the polynucleotide of SEQ ID NO: 419 encoding the variable heavy chain region of SEQ ID NO: 411; the poiinucleotides encoding the complementarity determining regions (SEQ ID NO: 420; SEQ ID NO: 421 and SEQ ID NO: 422) of the variable light chain region of SEQ ID NO: 410; and the polyjiucleotides encoding the complementarity determining regions (SEQ ID NO: 423; SEQ ID NO: 424 and SEQ ID NO: 425) of the variable heavy chain region of

332

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY SEQ ID NO: 411.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGGCCAGTGAGGACATTGAAAGCTATCTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGAATCTGGGGTCTCATCG

CGGTTC7VAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCC

GATGCTGCCACTTACTATTGTCAATGCACTTATGGTATTATTAGTATTAGTGATGGTAATGCT (SEQ ID NO: 434)

NO: 427:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGTGTCTGGATTCTCCCTCAGTAGCTACTTCATGACCTGGGTCCGCCAGGCTCCAGGGGAG

GGGCTGGAATACATCGGATTCATGAATACTGGTGATAACGCATACTACGCGAGCTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACC

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INDUSTRIAL

GAGGACACGGCCACCTATTTCTGTGCCAGGGTTCTTGTTGTTGCTTATGGAGCCTTTAACATC (SEQ ID NO: 435).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 436; SEQ ID NO: 437 and SEQ ID NO: 438 corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of SEQ ID NO: 426.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 439; SEQ ID NO: 440 and SEQ ID NO: 441 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td colspan="2">hypervariables) of the</td><td>sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody that

334

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INDUSTRIAL

possesses an IL-6 binding specificity comprising, or alternatively consisting of, one, two, three or more, including all of the following polynucleotides encoding antibody fragments: the polynucleotide of SEQ ID NO: 434 encoding the region variable light chain of SEQ ID NO: 426, the polynucleotide of SEQ ID NO: 435 encoding the variable heavy chain region of SEQ ID NO: 427; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 436; SEQ ID NO: 437 and SEQ ID NO: 438) of the variable light chain region of SEQ ID NO: 426; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 439; SEQ ID NO: 440 and SEQ ID NO: 441) of the variable heavy chain region of SEQ ID NO: 427.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGAACCTGTGGGAGGCACA

GTCAGCATCAGTTGCCAGTCCAGTAAGAGTGTTATGAATAACAACTACTTAGCCTGGTATCAG

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGCATCTGGGGTC

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MEXICAN INSTITUTE OF LA PRORIED. 'Ü

INDUSTRIAL

CCATCACGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAG

TGTGACGATGCTGCCACTTACTACTGTCAAGGCGGTTATACTGGTTATAGTGATCATGGGACT (SEQ ID NO: 450)

In. Another embodiment of the invention, the polynucleotides of the invention comprise or, alternatively, consist of the following polynucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID

NO: 443:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCAAGCCTGACGAAACCCTGACACTCACCTGC

ACAGTCTCTGGAATCGACCTCAGTAGCTATCCAATGAACTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGATTCATTAATACTGGTGGTACCATAGTCTACGCGAGCTGGGCAAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAGGCAGTTATGTTTCATCTGGTTATGCCTACTAT

TTTAATGTC (SEQ ID NO: 451).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 452; SEQ ID NO: 453 and SEQ ID NO: 454 corresponding to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 442.

In a further embodiment of the invention, the

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INDUSTRIAL

NO: 457 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 443.

The invention also contemplates polynucleotide sequences that include one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 450 encoding the variable light chain region of SEQ ID NO: 442, the polynucleotide of SEQ ID NO: 451 encoding the variable heavy chain region of SEQ ID NO: 443; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 452; SEQ ID NO: 453 and SEQ ID NO: 454) of the variable light chain region of SEQ ID NO:

442; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 455; SEQ ID NO:

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INDUSTRIAL

456 and SEQ ID NO: 457) of the variable heavy chain region of SEQ ID NO: 443.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACA

GTCAGCATCAGTTGCCAGTCCAGTCAGAGTGTTTATAATAACAACTGGTTATCCTGGTTTCAG

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTC

CCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAG

TGTGACGATGTTGCCACTTACTACTGTGCGGGCGGTTATCTTGATAGTGTTATT (SEQ ID

NO: 466)

NO: 459:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGTCTCTGGATTCTCCCTCAGTACCTATTCAATAAACTGGGTCCGCCAGGCTCCAGGGAAG

GGCCTGGAATGGATCGGAATCATTGCTAATAGTGGTACCACATTCTACGCGAACTGGGCGAAA

338 ¿Μ

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INDUSTRIAL

GGCCGATTCACCGTCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAGAGAGTGGAATGTACAATGAATATGGTAAATTT

AACATC (SEQ ID NO: 467).

NO: 470 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 458.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 471; SEQ ID NO: 472 and SEQ ID NO: 473 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td>heavy chain</td><td>variable of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, the

339

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INSTITUTO MSXICA »« J 'í

FROM PROPERTY 'J' j

INDUSTRIAL polynucleotides encoding fragments of the antibody possessing specificity for binding to IL-6 comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding antibody fragments: the polynucleotide of SEQ ID NO: 466 encoding the variable light chain region of SEQ ID NO: 458, the polynucleotide of SEQ ID NO: 467 encoding the variable heavy chain region of SEQ ID NO: 459; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 468; SEQ ID NO: 469 and SEQ ID NO: 470) of the variable light chain region of SEQ ID NO: 458; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 471; SEQ ID NO:

472 and SEQ ID NO: 473) of the variable heavy chain region of SEQ ID NO: 459.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTCTGATATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAATTGCCAGGCCAGTGAGAACATTTATAGCTTTTTGGCCTGGTATCAGCAGAAA

340

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INDUSTRMl

CCAGGGCAGCCTCCCAAGCTCCTGATCTTCAAGGCTTCCACTCTGGCATCTGGGGTCTCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGAC

GATGCTGCCACTTACTACTGTCAACAGGGTGCTACTGTGTATGATATTGATAATAAT (SEQ

ID NO: 482)

NO: 475:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACAGTTTCTGGAATCGACCTCAGTGCCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGGAG

GGGCTGGAATGGATCACAATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAA

GGCCGATTCACCGTCTCCAAAACCTCGACCGCGATGGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAGATGCAGAAAGTAGTAAGAATGCTTATTGGGGC

TACTTTAACGTC (SEQ ID NO: 483).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 484; SEQ ID NO: 485 and SEQ ID NO: 486 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 474.

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INDUSTRIAL

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 487; SEQ ID NO: 488 and SEQ ID NO: 489 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 482 encoding the variable light chain region of SEQ ID NO: 474, the polynucleotide of SEQ ID NO: 483 encoding the variable heavy chain region of SEQ ID NO: 475; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 484; SEQ ID NO: 485 and SEQ ID NO: 486) of the variable light chain region of SEQ ID NO: 474; and the polynucleotides that encode the regions

342

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Complementarity Determinants (SEQ ID NO: 487; SEQ ID NO:

wwnnBwiiTw.iu »an - <n 11 1

488 and SEQ ID NO: 489) of the variable heavy chain region of SEQ ID NO: 475.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTCTGATATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAATTGCCAGGCCAGTGAGAACATTTATAGCTTTTTGGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTTCAGGGCTTCCACTCTGGCATCTGGGGTCTCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGAC

GATGCTGCCACTTACTACTGTCAACAGGGTGCTACTGTGTATGATATTGATAATAAT (SEQ

ID NO: 498)

NO: 491:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACAGTTTCTGGAATCGACCTCAGTGCCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGGAG

343

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J-4. -lL MEXICAN INSTITUTE OF INDUSTPUAJ PROPERTY.

GGGCTGGAATGGATCACAATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAA

GGCCGATTCACCGTCTCCAAAACCTCGACCGCGATGGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAGATGCAGAAAGTAGTAAGAATGCTTATTGGGGC

TACTTTAACGTC (SEQ ID NO: 499).

NO: 502 corresponding to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 490.

NO: 505 that correspond to polynucleotides that encode the complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td colspan="4">of the variable heavy chain sequence of</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

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described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 498 encoding the variable light chain region of SEQ ID NO: 490, the polynucleotide of SEQ ID NO: 499 encoding the variable heavy chain region of SEQ ID NO: 4 91; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 500; SEQ ID NO: 501 and SEQ ID NO: 502) of the variable light chain region of SEQ ID NO: 490; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 503; SEQ ID NO:

504 and SEQ ID NO: 505) of the variable heavy chain region of SEQ ID NO: 491.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCCATTGAAATGACCCAGACTCCATCCCCCGTGTCTGCCGCTGTGGGAGGCACA

345

GTCACCATCAATTGCCAGGCCAGTGAGAGTGTTTTTAATAATATGTTATCCTGGTATCAGCAG aaaccagggcactctcctaagctcctgatctatgatgcatccgatctggcatctggggtccca

TCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGTGGCGTGGAGTGT

GACGATGCTGCCACTTACTATTGTGCAGGGTATAAAAGTGATAGTAATGATGGCGATAATGTT (SEQ ID NO: 514)

NO: 507:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACAGTCTCTGGATTCTCCCTCAACAGGAATTCAATAACCTGGGTCCGCCAGGCTCCAGGGGAG

GGGCTGGAATGGATCGGAATCATTACTGGTAGTGGTAGAACGTACTACGCGAACTGGGCAAAA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGAGGCCATCCTGGTCTTGGTAGTGGTAACATC (SEQ ID NO: 515).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 516; SEQ ID NO: 517 and SEQ ID NO: 518 which correspond to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

346

SEQ ID NO: 506.

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In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 519; SEQ ID NO: 520 and SEQ ID NO: 521 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td>of the</td><td>sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td colspan="2">contemplate sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 514 encoding the variable light chain region of SEQ ID NO: 506, the polynucleotide of SEQ ID NO: 515 encoding the variable heavy chain region of SEQ ID NO: 507; polynucleotides that encode complementarity determining regions (SEQ ID NO: 516; SEQ ID NO: 517 and SEQ ID NO:

518) of the variable light chain region of SEQ ID NO:

347

506; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 519; SEQ ID NO:

520 and SEQ ID NO: 521) of the variable heavy chain region of SEQ ID NO: 507.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCACATTTGCGCAAGTGCTGACCCAGACTGCATCGTCCGTGTCTGCAGCTGTGGGAGGCACA

GTCACCATCAATTGCCAGTCCAGTCAGAGTGTTTATAATAACTACTTATCCTGGTATCAGCAG

AAACCAGGGCAGCCTCCCAAGCTCCTGATCTATACTGCATCCAGCCTGGCATCTGGGGTCCCA

TCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGAAGTGCAGTGT

GACGATGCTGCCACTTACTACTGTCAAGGCTATTATAGTGGTCCTATAATTACT (SEQ ID

NO: 530)

NO: 523:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

348

ACAGCCTCTGGATTCTCCCTCAATAACTACTACATACAATGGGTCCGCCAGGCTCCAGGGGAG

GGGCTGGAATGGATCGGGATCATTTATGCTGGTGGTAGCGCATACTACGCGACCTGGGCAAAC

GGCCGATTCACCATCGCCAAAACCTCGTCGACCACGGTGGATCTGAAGATGACCAGTCTGACA

ACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGACATTTGATGGTTATGAGTTG (SEQ

ID NO: 531).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more poiinucleotide sequences of SEQ ID NO 532; SEQ ID NO: 533 and SEQ ID NO: 534 corresponding to poiinucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 522.

In a further embodiment of the invention, the poiinucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more poiinucleotide sequences of SEQ ID NO 535; SEQ ID NO: 536 and SEQ ID

NO: 537 that correspond to poiinucleotides that encode the complementarity determining regions (CDRs, or regions

<td>hypervariables) of the</td><td>sequence</td><td>chain</td><td>heavy</td><td>variable of</td><td>the</td>

<td>The invention</td><td>too</td><td colspan="3">contemplate sequences</td><td>of</td>

<td>25 poiinucleotides that</td><td>include</td><td>one or</td><td>plus</td><td>sequences</td><td>of</td>

349

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MEXICAN INSTITUTE OF PROPERTY / .t)

INDUSTRIAL polynucleotides encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-β binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 530 encoding the variable light chain region of SEQ ID NO: 522, the polynucleotide of SEQ ID NO: 531 encoding the variable heavy chain region of SEQ ID NO: 523; polynucleotides encoding the complementarity determining regions (SEQ ID NO: 532; SEQ ID NO: 533 and SEQ ID NO:

534) of the variable light chain region of SEQ ID NO:

522; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 535; SEQ ID NO:

536 and SEQ ID NO: 537) of the variable heavy chain region of SEQ ID NO: 523.

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

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GCCACATTTGCCCAAGTGCTGACCCAGACTCCATCCCCTGTGTCTGTCCCTGTGGGAGACACA

GTCACCATCAGTTGCCAGTCCAGTGAGAGCGTTTATAGTAATAACCTCTTATCCTGGTATCAG

CAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAGGGCATCCAATCTGGCATCTGGTGTC

CCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGCACAG

TGTGACGATGCTGCCACTTACTACTGTCAAGGCTATTATAGTGGTGTCATTAATAGT (SEQ

ID NO: 546)

NO: 539:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGC

ACAGTGTCTGGATTCTCCCTCAGTAGCTACTTCATGAGCTGGGTOCGCCAGGCTCCAGGGGAG

GGGCTGGAATACATCGGATTCATTAATCCTGGTGGTAGCGCATACTACGCGAGCTGGGCGAGT

GGCCGACTCACCATCTCCAAAACCTCGACCACGGTAGATCTGAAAATCACCAGTCCGACAACC

GAGGACACGGCCACCTATTTCTGTGCCAGGATTCTTATTGTTTCTTATGGAGCCTTTACCATC (SEQ ID NO: 547).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 548; SEQ ID NO: 549 and SEQ ID NO: 550 that correspond to polynucleotides that encode complementarity determining regions (CDRs, or regions

351

MEXICAN INSTITUTE OF PRO hypervariables) of the 'Wriable' light chain sequence

SEQ ID NO: 538

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 551; SEQ ID NO: 552 and SEQ ID NO: 553 corresponding to polynucleotides that encode complementarity determining regions (CDRs, or regions

<td>hypervariables)</td><td colspan="2">of the sequence of</td><td colspan="2">variable heavy chain</td><td>the</td>

<td colspan="2">The invention</td><td>too</td><td>behold</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td colspan="2">include one or more</td><td>sequences</td><td>of</td>

<td>polynucleotides</td><td>than</td><td>encode</td><td>fragments</td><td colspan="2">antibody</td>

described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 546 encoding the variable light chain region of SEQ ID NO: 538, the polynucleotide of SEQ ID NO: 547 encoding the variable heavy chain region of SEQ ID NO: 539; polynucleotides that encode complementarity determining regions (SEQ ID NO: 548; SEQ ID NO: 549 and SEQ ID NO:

352

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550) of the variable light chain region of SEQ ID NO: 538; and polynucleotides encoding the complementarity determining regions (SEQ ID NO: 551; SEQ ID NO:

552 and SEQ ID NO: 553) of the variable heavy chain region of SEQ ID NO: 539.

ATGGACACGAGGGCOCCCACTOAGCTGCTGGGGCTOCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCOTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGGCCACTGAGAGCATTGGCAATGAGTTATCCTGGTATCAGCAGAAA

CCAGGGCAGGCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACCGGCGTGGAGTGTGAT

GATGCTGCCACTTACTACTGTCAACAGGGTTATAGTAGTGCTAATATTGATAATGCT (SEQ

ID NO: 562)

NO: 555:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

353

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACCGTCTCTGGATTCTCCCTCAGTAAGTACTACATGAGCTGGGTCCGCCAGGCTCCAGAGAAG

GGGCTGAAATACATCGGATACATTGATAGTACTACTGTTAATACATACTACGCGACCTGGGCG

AGAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGTkAGATCACCAGTCCGACA

AGTGAGGACACGGCCACCTATTTCTGTGCCAGAGGAAGTACTTATTTTACTGATGGAGGCCAT

CGGTTGGATCTC (SEQ ID NO: 5 63).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 564; SEQ ID NO: 565 and SEQ ID NO: 566 which correspond to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 554.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 567; SEQ ID NO: 568 and SEQ ID NO: 569 which correspond to polynucleotides that encode the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 555.

The invention also contemplates sequences of

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INDUS ÍR1AL

polynucleotides that include one or more polynucleotide sequences encoding antibody fragments described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 562 encoding the variable light chain region of SEQ ID NO: 554, the polynucleotide of SEQ ID NO: 563 encoding the variable heavy chain region of SEQ ID NO: 555; polynucleotides that encode complementarity determining regions (SEQ ID NO: 564; SEQ ID NO: 565 and SEQ ID NO:

566) of the variable light chain region of SEQ ID NO:

554; and polynucleotides that encode complementarity determining regions (SEQ ID NO: 567; SEQ ID NO:

568 and SEQ ID NO: 569) of the variable heavy chain region of SEQ ID NO: 555.

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INDUSTRIAL

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGT

GCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACA

GTCACCATCAAGTGCCAGGCCACTGAGAGCATTGGCAATGAGTTATCCTGGTATCAGCAGAAA

CCAGGGCAGGCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACCGGCGTGGAGTGTGAT

GATGCTGCCACTTACTACTGTCAACAGGGTTATAGTAGTGCTAATATTGATAATGCT (SEQ

ID NO: 578)

NO: 571:

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGT

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTAACGCCTGGGACACCCCTCGACACTCACCTGC

ACAGTCTCTGGATTCTCCCTCAGTACCTACAACATGGGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGAAGTATTACTATTGATGGTCGCACATACTACGCGAGCTGGGCGAAA

GGCCGATTCACCGTCTCCAAAAGCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACC

GGGGACACGGCCACCTATTTCTGTGCCAGGATTCTTATTGTTTCTTATGGGGCCTTTACCATC (SEQ ID NO: 579).

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 580; SEQ ID NO: 581 and SEQ ID NO: 582 corresponding to polynucleotides encoding the

356 , 1 f A

MEXICAN in¿t: tl

£. '£ THE PROPERTY,, INDUilkwU. _ Complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of the

SEQ ID NO: 570.

In a further embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively, consist of one or more polynucleotide sequences of SEQ ID NO 583; SEQ ID NO: 584 and SEQ ID NO: 585 which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of the

SEQ ID NO: 571

The invention also contemplates polynucleotides that include one or more polynucleotides encoding fragments of antibody sequence sequences described herein. In one embodiment of the invention, polynucleotides encoding fragments of the antibody possessing IL-6 binding specificity comprise, or alternatively consist of, one, two, three, or more, including all of the following polynucleotides encoding fragments of Antibody: the polynucleotide of SEQ ID NO: 578 encoding the variable light chain region of SEQ ID NO: 570, the polynucleotide of SEQ ID NO: 579 encoding the variable heavy chain region of SEQ ID NO: 571; the polynucleotides that encode the determining regions of

357

institute μ ·. ·?; κ; λ; · '> D £ U \ ΡΗΟΓΤΓΓχ -.'-, υ INDVü l il'AL

Complementarity (SEQ ID NO: 580; SEQ ID NO: 581 and SEQ ID NO: 582) of the variable light chain region of SEQ ID NO: 570; and the poiinucleotides encoding the complementarity determining regions (SEQ ID NO: 583; SEQ ID NO:

584 and SEQ ID NO: 585) of the variable heavy chain region of SEQ ID NO: 571.

In another embodiment of the invention, the poiinucleotides of the invention further comprise or, alternatively, consist of the following poiinucleotide sequence encoding the kappa constant light chain sequence of SEQ ID NO: 586:

GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGA

ACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAG

GTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGAC

AGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTC

TACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGA

GAGTGT (SEQ ID NO: 587).

In another embodiment of the invention, the poiinucleotides of the invention additionally comprise, or alternatively consist of, the following poiinucleotide sequence encoding the gamma-1 constant heavy chain polypeptide sequence of SEQ ID NO: 588:

GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCT

GGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCG

TGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGA

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CTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC

TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGT

GACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTC

CTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTG

GTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAG

GTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGC

GTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAAC

AAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCA

CAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGC

CTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAG

AACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAG

CTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAG

GCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA (SEQ ID NO:

589).

In another embodiment of the invention, the polynucleotides of the invention additionally comprise, or alternatively consist of, the following polynucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID NO: 700:

GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGA

CTCTCCTGTGCAGCCTCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGTCAGGCT

CCAGGGAAGGGGCTGGAGTGGGTCGGCATCATCTATGGTAGTGATGAAACCGCCTACGCTACC

TCCGCTATAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATG

AACAGCCTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGATGATAGTAGTGACTGG

GATGCAAAGTTCAACTTGTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC (SEQ ID NO:

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MEXICAN INSTITUTE 'tÓ r> ria PUfVPlTUAD V

700) .

In another embodiment of the invention, polynucleotides of.

L — βίβλοι *. ·? - - .'τΓ. ^ ,. ντ trz.-ygJniiT »** ·> * · *» the invention additionally comprises, or alternatively consists of the following polynucleotide sequence encoding the variable light chain polypeptide sequence of SEQ ID NO: 723:

GCTATCCAGATGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTC

ACCATCACTTGCCAGGCCAGTCAGAGCATTAACAATGAGTTATCCTGGTATCAGCAGAAACCA

GGGAAAGCCCCTAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCCCATCAAGG

TTCAGCGGCAGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGAT

TTTGCAACTTATTACTGCCAACAGGGTTATAGTCTGAGGAACATTGATAATGCTTTCGGCGGA

GGGACCAAGGTGGAAATCAAACGT (SEQ ID NO: 723).

In one embodiment, the invention is directed to an isolated polynucleotide comprising a polynucleotide encoding an antibody V amino acid sequenceH anti-IL-6

<td colspan="4">selected from the</td><td>SEQ ID</td><td>NOT:</td><td colspan="2"> 3, 18, 19,</td><td> 22,</td><td> 38, 54, 70,</td><td> 86,</td>

708 or encodes a variant thereof where at least one flanking residue (FR residue) was replaced by an amino acid present at the corresponding position in a rabbit anti-IL-6 antibody VH polypeptide or a conservative amino acid substitution.

360

ΙΜΡΙϋ> ^

LNÜ .7, / 1 7 ί //> ·! 7 <> .a V, ü-cdí ji DL LA Ef.c ·· ;. :, \ D V. ~ U7 ~ J · ί®

INUSTICAL gjg ^

In another embodiment, the invention is directed to an isolated polynucleotide comprising the poiinucleotide sequence encoding an anti-IL-6 VL antibody amino acid sequence selected from SEQ ID NO: 2, 20,

<td colspan="2">or 709 or</td><td>encode a</td><td colspan="2">variant</td><td>of the</td><td colspan="2">same where to</td><td colspan="2">minus one</td>

<td colspan="2">residue</td><td>flanking</td><td colspan="2">(residue</td><td>FR)</td><td>It was</td><td>replaced</td><td>by</td><td>a</td>

amino acid present at the corresponding position in a rabbit anti-IL-6 antibody VL polypeptide or a conservative amino acid substitution.

In yet another embodiment, the invention is directed to one or more heterologous polynucleotides comprising a sequence encoding the polypeptides contained in SEQ ID NO: 2 and SEQ ID NO: 3; SEQ ID NO: 2 and SEQ ID NO: 18; SEQ ID NO: 2 and SEQ ID NO: 19;

SEQ ID NO: 20 and SEQ ID NO: 3; SEQ ID NO: 20 and SEQ ID NO: 18; SEQ ID

NO: 20 and SEQ ID NO: 19; SEQ ID NO: 21 and SEQ ID NO: 22; SEQ ID NO: 37 and SEQ ID NO: 38; SEQ ID NO: 53 and SEQ ID NO: 54; SEQ ID NO: 69 and SEQ ID NO: 70; SEQ ID NO: 85 and SEQ ID NO: 86; SEQ ID NO: 101 and SEQ ID NO: 102; SEQ ID NO: 101 and SEQ ID N0: 117; SEQ ID NO: 101 and SEQ ID

NO: 118; SEQ ID NO: 119 and SEQ ID NO: 102; SEQ ID NO: 119 and SEQ ID

NO: 117; SEQ ID NO: 119 and SEQ ID NO: 118; SEQ ID NO: 122 and SEQ ID

NO: 123; SEQ ID NO: 138 and SEQ ID NO: 139; SEQ ID NO: 154 and SEQ ID

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In another embodiment, the invention is directed to an isolated polynucleotide that expresses a polypeptide containing at least one CDR polypeptide that is derived from an anti-IL-6 antibody, where said expressed polypeptide binds only specifically to IL-6 or specifically binds to IL-6 when expressed in association with another polynucleotide sequence that expresses a polypeptide containing at least one CDR polypeptide that comes from an anti-IL-6 antibody, where said at least one CDR is selected from those contained in the V polypeptidesL or VH contained in SEQ ID NO: 3, 18, 19, 22, 38, 54, 70, 86, 102, 117, 118, 123, 139, 155, 171, 187,

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<td> 679,</td><td> 683,</td><td> 687,</td><td> 693,</td><td> 699,</td><td> 702,</td><td> 706,</td><td colspan="2">or [sic].</td><td colspan="3">The sequence of</td>

Example nucleic acid encoding the VH and VL polypeptides of SEQ ID NO: 657 and SEQ ID NO: 709 are comprised of SEQ ID NO: 700 and SEQ ID NO: 723, respectively

Host cells and vectors comprising such polynucleotides are also contemplated.

The invention further contemplates vectors comprising the polynucleotide sequences encoding the variable heavy and variable light chain polypeptide sequences, as well as the determining regions of

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OF PROPERTY Vc-&T—TAT: ',

INDUSTRIAL ^ »_ ίίϊ__Χ -> * individual complementarity (CDR or hypervariable regions) established herein, as well as host cells comprising such sequences. In an embodiment of the invention, the host cell is a yeast cell. In another embodiment of the invention, the yeast host cell belongs to the genus Pichia.

In some cases, more than one example of a polynucleotide encoding a given polypeptide sequence is provided, as summarized in Table 3.

Table 3. Multiple examples of polynucleotides encoding particular polypeptides.

<td>Polypeptide SEQ ID NO:</td><td>Examples of SEQ ID NO coding</td>

364

ΙΜ

365

In some cases, multiple sequence identifiers refer to the same polypeptide or polynucleotide sequence, as summarized in Table 4. References to these sequence identifiers are understood to be interchangeable, except where the context indicates otherwise.

Table 4. Repeated sequences. Each cell lists a

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group of repeated sequences included in the sequence listing.

<td>SEQ ID NO of sequences repeated</td>

367

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Some examples of modalities include polynucleotides that hybridize under conditions of moderate or high restriction hybridization to a polynucleotide having one of the examples of coding sequences cited in Table 1, and also include polynucleotides that hybridize under conditions of moderate or high restriction hybridization to a polynucleotide encoding the same polypeptide as a polynucleotide having one of the examples of coding sequences cited in Table 1, or a polypeptide encoded by any of the anterior polynucleotides.

The term "high restriction hybridization conditions" refers to conditions under which a probe hybridizes to its target subsequence, typically in a complex nucleic acid mixture but not to other sequences. The high restriction conditions depend on the sequences and will differ in different circumstances. Longer sequences hybridize specifically at higher temperatures. A comprehensive guide to nucleic acid hybridization is found in Tijssen, Techniques in Biochemistry and Molecular Biology — Hybridization with Nucleic Probes, OverView of principles of hybridization and the strategy of nucleic acid assays (1993). In general, the high restriction conditions are stated to be around 5-10 ° C less than the thermal melting point (Tm) for the specific sequence at pH and

369

ΡΙ

INSTITUTO MEXICANO 'DE LA PROI'ÍF.DaD • INDUSTRIAL defined ionic strength. Tm is the temperature (according to defined nucleic concentration, pH and ionic strength) at which 50% of the complementary probes to the target hybridize the target sequence in equilibrium (as the target sequences are present in excess, at Tm, 50 % of probes are in equilibrium). High restriction conditions will be those where the salt concentration is less than about 1.0 M sodium ion concentration, typically around 0.01 to 1.0 M sodium ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least around 30 ° C for short probes (eg, 10 to 50 nucleotides) and at least around 60 ° C for long probes (eg, greater than 50 nucleotides). High restriction conditions can also be achieved with the addition of destabilizing agents such as formamide. For selective or specific hybridization, a positive signal is at least twice background hybridization, optionally 10 times background hybridization. Examples of high restriction hybridization conditions can be as follows: 50% formamide, 5xSSC and 1% SDS, incubating at 42 ° C, or 5xSSC, 1% SDS, incubating at 65 ° C, with 0.2 wash xSSC, and 0.1% SDS at 65 ° C. Such hybridization and washing steps can be performed during, eg, 1, 2, 5, 10, 15, 30, 60; or more minutes.

Nucleic acids that do not hybridize to each other under high restriction conditions are still substantially

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related if the polypeptides they encode are substantially related. This occurs, for example, when a copy of a nucleic acid is created using the maximum codon degeneracy allowed by the genetic code. In these cases, nucleic acids typically hybridize under conditions of moderate restriction hybridization. Examples of moderate restriction hybridization conditions include hybridization in a buffer of 40% formamide, 1M NaCl, 1% SDS at 37 ° C and a wash in 1> <SSC at 45 ° C. Such hybridization and washing steps can be performed for, eg, 1, 2, 5, 10, 15, 30, 60, or more minutes. A positive hybridization is at least twice background hybridization. Those skilled in the art will quickly recognize that alternative hybridization and washing conditions can be used to provide similar restriction conditions.

Examples of modalities of heavy and light chain polypeptides and polynucleotides

This section describes examples of modalities of heavy and light chain polypeptides, as well as examples of polynucleotides that encode such polypeptides. These examples of polynucleotides are suitable for expression in the described Pichia expression system.

In certain embodiments, the present invention comprises polynucleotides possessing at least 70%, such as at least 75%, at least 80%, at least 85%, at least 90%, at least

371

91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the polynucleotides described in this application or that encode polypeptides described in this application or that hybridize to said polynucleotides under conditions of low restriction, moderate restriction or high restriction, preferably those that encode polypeptides (eg. an immunoglobulin heavy and light chain, a single chain antibody, an antibody fragment, etc.) that possess at least one of the biological activities set forth herein, including but not limited to specific binding to a polypeptide of IL-6. In another aspect, the invention comprises a composition comprising such a polynucleotide and / or a polypeptide encoded by such a polynucleotide. In yet another aspect, the invention comprises a method of treating an IL-6 associated disease or condition that can be prevented, treated or ameliorated with an IL-6 antagonist such as Abl (eg cachexia, cancer fatigue , arthritis, etc.) comprising administering a composition comprising such a polynucleotide and / or polypeptide.

In some preferred embodiments, a heavy chain polypeptide will comprise one or more CDR sequences of the heavy and / or light chain polypeptides described herein (including those contained in the heavy and light chain polypeptides described herein) and one or more

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Flanking region polypeptides described herein, including those depicted in Figures 2 and '34 -37 or * Table 1, and contained in the heavy and light chain polypeptide sequences described herein. In certain preferred embodiments, a heavy chain polypeptide will comprise one or more flanking region 4 sequences as depicted in Figures 2 and 34-37 or Table 1, or as contained in heavy or light chain polypeptides described herein. . In certain preferred embodiments, a light chain polypeptide will comprise one or more of the CDR sequences of the heavy and / or light chain polypeptides described herein (including those contained in the heavy and light chain polypeptides described herein). and one or more of the flanking region polypeptides described herein, including those depicted in Figures 2 and 34-37 or Table 1, and contained in the heavy and light chain polypeptide sequences described herein. In certain preferred embodiments, a light chain polypeptide will comprise one or more flanking region 4 sequences as depicted in Figures 2 and 34-37 or Table 1, or as contained in heavy or light chain polypeptides described herein. .

In any of the described modalities. Herein, certain described sequences may be

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substituted between them, unless the context indicates otherwise. The fact that particular sequences may be substituted for each other, where such events occurred, is understood to be illustrative and not limiting in nature, and such substitutions are also understood to be encompassed even though illustrative examples of substitutions were not described, for example, where one or more of the Abl light chain polypeptides is described, eg.

any of SEQ ID NO: 2, 20, 647, 651, 660, 666, 699,

702, 706, or 709, another Abl light chain polypeptide can be substituted unless the context indicates otherwise. Similarly, where one of the Abl heavy chain polypeptides is described, eg any of SEQ ID NO: 3, 18,

19, 652, 656, 657, 658, 661, 664, 665, 704, or 708, another Abl heavy chain polypeptide may be substituted unless the context indicates otherwise. Likewise, where one of the Abl light chain polynucleotides is described, eg. any of SEQ ID NO: 10, 662, 698, 701, or 705, another Abl light chain polynucleotide may be substituted unless the context indicates otherwise. Similarly, where one of the heavy chain polynucleotides is described

<td>Abl, for example</td><td>. any of SEQ ID NO: 11,</td><td> 663,</td><td> 700, 703,</td>

<td>or 707, other</td><td>heavy chain polynucleotide</td><td>of</td><td>Abl can</td>

<td>replace</td><td>unless the context indicates</td><td>the</td><td>contrary.</td>

<td>further</td><td>, it is understood that the description</td><td>of</td><td>any</td>

374 member of any of the following groups ^1 comprises substitution by any other ± group member, as follows: Ab2 light chain polypeptides (SEQ ID NO: 21 and

<td> 667) ;</td><td>Ab2 light chain polynucleotides (SEQ ID NO: 29 and</td>

<td> 669) ;</td><td>Ab2 heavy chain polypeptides (SEQ ID NO: 22 and</td>

<td> 668) ;</td><td>Ab2 heavy chain polynucleotides (SEQ ID NO: 30 and</td>

<td> 670) ;</td><td>Ab3 light chain polypeptides (SEQ ID NO: 37 and</td>

<td> 671) ;</td><td>Ab3 light chain polynucleotides (SEQ ID NO: 45 and</td>

<td> 673) ;</td><td>Ab3 heavy chain polypeptides (SEQ ID NO: 38 and</td>

<td> 672) ;</td><td>Ab3 heavy chain polynucleotides (SEQ ID NO: 46 and</td>

<td> 674) ;</td><td>Ab4 light chain polypeptides (SEQ ID NO: 53 and</td>

<td> 675) ;</td><td>Ab4 light chain polynucleotides (SEQ ID NO: 61 and</td>

<td> 677) ;</td><td>Ab4 heavy chain polypeptides (SEQ ID NO: 54 and</td>

<td> 676) ;</td><td>Ab4 heavy chain polynucleotides (SEQ ID NO: 62 and</td>

<td> 678) ;</td><td>Ab5 light chain polypeptides (SEQ ID NO: 69 and</td>

<td> 679) ;</td><td>Ab5 light chain polynucleotides (SEQ ID NO: 77 and</td>

<td> 681) ;</td><td>Ab5 heavy chain polypeptides (SEQ ID NO: 70 and</td>

<td> 680) ;</td><td>Ab5 heavy chain polynucleotides (SEQ ID NO: 78 and</td>

<td> 682) ;</td><td>Ab6 light chain polypeptides (SEQ ID NO: 85 and</td>

<td> 683) ;</td><td>Ab6 light chain polynucleotides (SEQ ID NO: 93 and</td>

<td> 685) ;</td><td>Ab6 heavy chain polypeptides (SEQ ID NO: 86 and</td>

<td> 684) ;</td><td>Ab6 heavy chain polynucleotides (SEQ ID NO: 94 and</td>

<td> 686) ;</td><td>Ab7 light chain polypeptides (SEQ ID NO: 101,</td>

119, 687, 693); Ab7 light chain polynucleotides (SEQ ID NO: 109 and 689); Ab7 heavy chain polypeptides (SEQ ID

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NO: 102, 117, 118, 688, 691, and 692); Ab7 heavy chain polynucleotides (SEQ ID NO: 110 and 690); Abl light chain CDR1 polynucleotides (SEQ ID NO: 12 and 694); Abl light chain CDR3 polynucleotides (SEQ ID NO: 14 and 695); Abl heavy chain CDR2 polynucleotides (SEQ ID NO: 16 and 696); and Abl heavy chain CDR3 polynucleotides (SEQ ID NO: 17 and 697).

Anti-IL-6 activity

As previously stated, IL-6 is a member of the cytokine family that promotes cellular responses through a receptor complex consisting of at least one subunit of the gpl30 signal transducer glycoprotein and the IL-receptor. 6 (IL-6R). IL-6R may also be present in soluble form (sIL-6R). IL-6 binds to IL-6R, which then dimerizes the gpl30 signal transducer receptor.

The anti-IL-6 antibodies of the invention, or IL-6 binding fragments thereof, are believed to be useful in exhibiting anti-IL-6 activity. In a non-limiting embodiment of the invention, the anti-IL-6 antibodies of the invention or the IL-6 binding fragments thereof, exhibit anti-IL-6 activity by binding to IL-6 which can be Soluble IL-6 or IL-6 expressed on the cell surface and / or can prevent or inhibit IL-6 binding to IL-6R and / or

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activation (dimerization) of the gpl30 signal transducing glycoprotein and the formation of IL-6 / IL-6R / gpl30 muitimers and the biological effects of any of the above. The anti-IL-6 antibodies in question may possess different antagonistic activities depending on where (i.e. epitope) the particular antibody binds to IL-6 and / or how it affects the formation of the above IL-6 complexes and / or muitimers and the biological effects thereof. As a consequence, different anti-IL-6 antibodies according to the invention, eg. they may be more suitable for preventing or treating conditions that involve the formation and accumulation of substantial soluble IL-6 such as rheumatoid arthritis while other antibodies may be favorable in treatments where the prevention of IL-6 / IL-6R / gpl30 or IL muitimers -6 / IL-6R / gpl30 is the desired therapeutic result. This can be determined in binding and other tests.

The anti-IL-6 activity of the anti-IL-6 antibody of the present invention and fragments thereof possessing IL-6 binding specificity may also be described by its binding strength or its affinity for IL-6. This can also affect its therapeutic properties. In one embodiment of the invention, the anti-IL-6 antibodies of the present invention and fragments thereof possessing IL-6 binding specificity bind to IL-6 with a dissociation constant (KD) less than or equal to 5xl0 ~7, ΙΟ '7, 5xl0-8, 10~8, 5x10 '

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MEXICAN INSTITUTE \

From LA PKO - '! £ Oa1') \ S' industrial V

10'9, 5xlO ~10, 10_1°, 5xl0 'or, 10'n, 5xl0-12, 10'12, 5xl0 '13

5xlCT14, 10 14, 5xl0 ~15 or 10-15. Preferably, anti-IL-6 antibodies and fragments thereof bind to IL-6 with a dissociation constant of less than or equal to 5xlO "10

In another embodiment of the invention, the anti-IL-6 activity of the anti-IL-6 antibodies of the present invention and fragments thereof possessing binding specificity for

IL-6, bind to IL-6 with a dissociation rate less than or equal to 10 '4 S '1, 5xl0-5 S '1, 10’5 S_1, 5xl0-6 S_1, 10’6 S '1, 5x10 '7 S1, or

10’7 S1. In one embodiment of the invention, the anti-IL-6 antibodies of the invention and fragments thereof possessing IL-6 binding specificity bind to a linear or conformational epitope of IL-6.

In a further embodiment of the invention, the anti-IL-6 activity of the anti-IL-6 antibodies of the present invention and fragments thereof which possess specificity for binding to IL-6, exhibit anti-IL-6 activity by improvement or reduction of symptoms of, or alternatively treating or preventing diseases and disorders associated with IL6. Non-limiting examples of IL-6 associated diseases and disorders are set forth below. As noted, fatigue related to cancer, cachexia, and rheumatoid arthritis are preferred indications for the anti-IL-6 antibodies in question.

In another embodiment of the invention, anti378 antibodies

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IL-6 described herein, or the IL-6 binding fragments thereof, do not possess binding specificity for IL-6R or the gp-130 signal transducing glycoprotein.

Analysis and isolation of B lymphocytes

In one embodiment, the present invention provides methods of isolating a clonal population of antigen-specific B lymphocytes that can be used to isolate at least one antigen-specific cell. As described and exemplified below, these methods contain a series of cultivation and selection steps that can be used separately, in combination, sequentially, repetitively, or periodically. These methods are preferably used to isolate at least one antigen-specific cell that can be used to produce a monoclonal antibody that is specific for a desired antibody or a nucleic acid sequence that corresponds to such an antibody.

In one embodiment, the present invention provides a method comprising the steps of:

to. preparing a cell population comprising at least one antigen-specific B lymphocyte;

b. enriching the cell population, eg, by chromatography, to form an enriched cell population comprising at least one antigen-specific B lymphocyte;

c. isolate the simple B lymphocyte from the population

379 enriched with B lymphocytes; and

d. determine if the simple B lymphocyte produces an antigen-specific antibody.

In another embodiment, the present invention provides an improvement to a method of isolating a single antibody-producing B lymphocyte, this improvement comprises enriching the population of B lymphocytes obtained from a host that was immunized or naturally exposed to an antigen, where the enrichment stage precedes any selection stage, it comprises at least one culture step and results in a clonal population of B lymphocytes that produce a single monoclonal antibody specific for said antigen.

Throughout this application, a clonal population of B lymphocytes refers to a population of B lymphocytes that only secretes a single antibody specific for a desired antigen. That is, these cells produce only one type of monoclonal antibody specific for the desired antigen.

In the present application, enriching a cell population means increasing the frequency of desired cells, typically antigen-specific cells, contained in a mixed cell population, eg, an isolate containing B lymphocytes from a host that is immunized against a desired antigen. Thus, an enriched cell population comprises a cell population possessing a higher frequency of antigen-specific cells such as

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result of an enrichment step, but this cell population can contain and produce different antibodies.

The general term cell population encompasses pre-enrichment and post-enrichment cell populations, keeping in mind that when multiple enrichment steps are performed, a cell population can be pre-enrichment or post-enrichment. For example, in one embodiment, the present invention provides a method:

to. which cultivates a cell population from an immunized host to obtain a cultured cell population;

b. that it creates at least one single-cell suspension from the cultured cell population;

c. which enriches at least one unicellular suspension to form a first enriched cell population;

d. which enriches the first enriched cell population to form a second enriched cell population;

and. which enriches the second enriched cell population to form a third enriched cell population; and

F. that selects an antibody produced by an antigen-specific cell from the third enriched cell population.

Each cell population can be used directly in the next stage or can be partially or totally frozen for short or long-term storage or for stages

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Later MEXICAN INSTITUTE OF PROPERTY. In addition, cells of a surrounding Cell pot can be individually suspended to provide single cell suspensions. The unicellular suspension can be enriched to such an extent that a unicellular suspension serves as the pre-enrichment cell population. Then, one or more of the antigen-specific unicellular suspensions together form the enriched cell population;

antigen-specific unicellular suspensions can be pooled, eg, plated again for further analysis and / or antibody production.

In one embodiment, the present invention provides a method of enriching a cell population to give an enriched cell population that has an antigen-specific cell frequency of between about 50% and

100% or increments thereof.

Preferably, the enriched cell population has an antigen-specific cell frequency greater than or equal to about 50%, 60%, 70%, 75%, 80%, 90%, 95%, 99%, or 100%.

In another embodiment, the present invention provides a method of enriching a cell population by increasing the frequency of antigen-specific cells at least 2-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or increases thereof.

Throughout this application, the term increment is used to define a numerical value to a greater or lesser degree of

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precision, for example, as close as possible to 10, 1, 0.1, 0.01, etc. The increment can be rounded to any measurable degree of precision and the increment must not be rounded to the same degree of precision at both ends of a range. For example, the range from 1 to 100 or increments thereof includes ranges such as 2 to 80, 5 to 50, and 0.4 to 98. When a range has no fixed limit, eg, a range less than 100, increments of It means increments between 100 and the measurable limit. For example, less than 100 or increments thereof means 0 to 100 or increments thereof unless the property, eg, temperature, is not limited by 0.

The specificity for antigen can be measured with respect to any antigen. The antigen can be any substance to which an antibody can bind including, but not limited to, peptides, proteins or fragments thereof, carbohydrates; organic and inorganic molecules; receptors produced by animal cells, bacterial cells, and viruses; enzymes; agonists and antagonists of biological pathways;

hormones; and cytokines. Examples of antigens include but are not limited to IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-18,

IFN-α, IFN-γ, BAFF, CXCL13, IP-10, VEGF, EPO, EGF, HRG, hepatocyte growth factor (HGF) and Hepcidin. Preferred antigens include IL-6, IL-13, TNF-oi, VEGF-α, hepatocyte growth factor (HGF), and Hepcidin. In a method that uses more than one enrichment step, the antigen used

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in each enrichment stage it can be the same or different in each stage. Multiple enrichment steps with the same antigen can yield a large and / or diverse population of antigen-specific cells; the multiple stages of enrichment with different antigens can give an enriched cell population with cross specificity for the different antigens.

Enrichment of a cell population can be carried out by any cell selection means known in the art for the isolation of antigen-specific cells. For example, a cell population can be enriched by chromatographic techniques, eg, Miltenyi bead technology or magnetic beads. The beads can bind directly or indirectly to the antigen of interest. In a preferred embodiment, the method of enriching a cell population includes at least one chromatographic enrichment step.

A cell population can also be enriched by any antigen specificity assay technique known in the art, eg, an ELISA assay or halogen assay. ELISA assays include but are not limited to selective antigen immobilization (eg, capture of biotinylated antigen by streptavidin, avidin, or neutravidin coated plates), plaque coating for non-specific antigen and through a concentration strategy of

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antigen (eg, selective antigen capture followed by addition of a binding partner to generate a heteromeric protein-antigen complex). The antigen can be attached directly or indirectly to a matrix or solid support, eg.

a column. A halogen assay comprises contacting cells with beads loaded with antigen and the host-specific labeled anti-host antibody used for the cultivation of B lymphocytes. The label may be, eg, a fluorophore. In one embodiment, at least one enrichment step of the assay is performed on at least one single-cell suspension. In another embodiment, the method of enriching a cell population includes at least one chromatographic enrichment step and at least one assay enrichment step.

Methods of enriching a cell population by size or density are known in the art. See, eg.

US Patents 5,627,052. These steps can be used in the present method additionally to enrich the cell population by antigen specificity.

The cell populations of the present invention contain at least one cell capable of recognizing an antigen. Cells that recognize antigens include, but are not limited to, B lymphocytes, plasma cells, and progenies thereof.

In one embodiment, the present invention provides a clonal cell population containing a single type of

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MEXICAN INSTITUTE OF THE. INDUSTRIAL PROPERTY antigen-specific B lymphocytes, that is, the cell population produces a single monoclonal antibody specific for a desired antigen.

In such an embodiment, the B lymphocyte antigen-specific clonal population is believed to consist predominantly of antigen-specific cells, antibody-secreting cells obtained by the new culture, and the selection protocol provided herein. Accordingly, the present invention also provides methods for obtaining an enriched cell population containing at least one antigen-specific cell and secreting an antibody. In one embodiment, the present invention provides an enriched cell population containing about 50% to about 100%, or increments thereof, or more than or equal to about 60%, 70%, 80%, 90%, or

100% of cells specific for antigen and that secrete an antibody.

In one embodiment, the present invention provides a method of isolating a single B lymphocyte by enriching a cell population obtained from a host prior to the selection steps, eg, selecting a particular B lymphocyte from a cell population and / or selecting an antibody produced by a particular cell. The enrichment stage can be carried out in one, two, three or more stages. In one embodiment, a single B lymphocyte is isolated to

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from an enriched cell population before confirming whether the simple B lymphocyte secretes an antibody with antigen specificity and / or a desired property.

In one embodiment, a method for enriching a cell population is used in a method for the production and / or selection of antibodies. Therefore, the present invention provides a method that comprises enriching a cell population before selecting an antibody. The method may include the steps of: preparing a cell population comprising at least one antigen-specific cell, enriching the cell population by isolating at least one antigen-specific cell to form an enriched cell population, and inducing antibody production from at least one cell specific for antigen. In a preferred embodiment, the enriched cell population contains more than one antigen-specific cell. In one embodiment, each antigen-specific cell in the enriched population is cultured under conditions that provide a clonal population of antigen-specific B lymphocytes before isolating a cell that produces antibodies thereto and / or producing an antibody using said B lymphocyte or a nucleic acid sequence that corresponds to that antibody. Unlike previous techniques where antibodies are produced from a cell population with a low frequency of antigen-specific cells, the present invention allows the

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selection of antibodies from a high frequency of antigen-specific cells. Because an enrichment step is used prior to antibody selection, most cells, preferably, and virtually all cells, used for antibody production are antigen-specific. By producing antibodies from a population of cells with an increased frequency of antigen specificity, the amount and variety of antibodies increases.

In the antibody selection methods of the present invention, an antibody is preferably selected after an enrichment step and a culture step that results in a clonal population of antigen-specific B lymphocytes. The methods may further comprise a step of determining the sequence of a selected antibody or portions thereof from one or more isolated antigen-specific cells. Any method known in the art can be employed to determine sequences and this may include determining the sequence of the heavy chain, light chain, variable region (s) and / or complementary region determining region (s) (CDR).

In addition to the enrichment step, the antibody selection method may also include one or more steps of analysis of the cell population for antigen recognition and / or antibody functionality. For example,

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I

Desired antibodies can have specific structural characteristics, such as binding to a particular epitope or mimicry of a particular structure; antagonist or agonist activity or neutralizing activity, eg, inhibit the binding between the antigen and a ligand. In one embodiment, analysis of antibody functionality depends on the ligand. Analysis of antibody functionality includes, but is not limited to, an in vitro protein-protein interaction assay that recreates the natural interaction of the antigen ligand with the recombinant receptor protein; and a ligand-dependent, cell-based response that is easily monitored (eg, proliferation response). In one embodiment, the method for antibody selection includes a step of analyzing the cell population for the functionality of the antibody by measuring the inhibitory concentration (IC50). In one embodiment, at least one of the antigen-specific isolated cells produces an antibody with an IC50 less than about 100, 50, 30, 25, 10 pg / mL or increases thereof.

In addition to the enrichment step, the antibody selection method may also include one or more steps of analyzing the cell population to determine the binding strength of the antibody. Antibody binding strength can be measured by any method known in the art (eg Biacore ™). In one embodiment, at least one of the cells

389

Isolated and antigen-specific produces an antibody that has a high affinity to the antigen, eg, a dissociation constant (Kd) of less than about 5χ10-1θ Ml, preferably around lxlO-13 and 5xlO ~10, lxlO-12 and lxlO-10, lxlO-12 and 7.5xl0-11, lxlO '11 and 2xl0-11, about 1.5xl0 ~n or less, or increases thereof. In this embodiment, the antibodies are said to be of mature affinity.

In a preferred embodiment, the affinity of the antibodies can be compared to or is greater than the affinity of either Panorex® (edrecolomab), Rituxan® (rituximab), Herceptin® (traztuzumab), Mylotarg® (gentuzumab), Campath® ( alemtuzumab),

Zevalin ™ (ibritumomab), Erbitux ™ (cetuximab), Avastin ™ (bevicizumab), Raptiva ™ (efalizumab), Remicade® (infliximab), Humira ™ (adalimumab), and Xolair ™ (omalizumab). Preferably, the affinity of the antibodies can be compared to or is greater than the affinity of Humira ™. The affinity of an antibody can also be increased by known affinity maturation techniques. In one embodiment, at least one cell population is analyzed to determine at least one, preferably both, the functionality of the antibody and the binding strength of the antibody.

In addition to the enrichment step, the antibody selection method may also include one or more steps of analyzing the cell population to determine antibody sequence homology, especially homology in

390

humans. In one embodiment, at least one of the isolated, antigen-specific cells produces an antibody that has homology with a human antibody of about 50% to about 100%, or increases in these, or more than about 60%, 70% , 80%, 85%, 90%, or 95% homologous. Antibodies can be humanized to increase homology to a human sequence by techniques known in the art such as CDR grafting or selectivity determining residue (SDR) grafting.

In another embodiment, the present invention also provides the antibodies themselves according to any of the modalities described above for IC50, Kd and / or homology.

The B lymphocyte selection protocol described herein has several intrinsic advantages over other methods for obtaining B lymphocytes that secrete monoclonal antibodies and antibodies specific for the desired target antigens. These benefits include, but are not limited to, the following:

First, it has been discovered that when these selection procedures are used with a desired antigen such as IL-6 or TNF-α, these methods reproducibly result in antigen-specific B lymphocytes capable of generating what appears to be be a substantially integral complement of the antibodies, that is, antibodies that bind to the different epitopes of the antigen.

391

Without limiting itself to theory, it is hypothesized that the

<td>complement</td><td>integral is</td><td></td><td>attributable</td><td>to the stage</td><td>of</td>

<td colspan="3">antigen enrichment</td><td>That</td><td>performed before</td><td>the</td>

<td>Recovery</td><td>lymphocyte</td><td>B</td><td>initial.</td><td colspan="2">Also, this advantage</td>

<td>5 allows the</td><td>insulation and</td><td>the</td><td colspan="2">antibody selection</td><td>with</td>

different properties as these properties can vary depending on the epitopic specificity of the particular antibody.

Second, the B lymphocyte selection protocol has been found to reproducibly produce a clonal B lymphocyte culture containing a single B lymphocyte or its progeny, secreting a single monoclonal antibody that generally binds the desired antigen with a relatively high binding affinity, that is, picomolar antigen binding affinities or better. Instead, the above methods of antibody selection tend to produce few high-affinity antibodies and therefore require extensive testing procedures to isolate an antibody with therapeutic potential. Without limiting itself to theory, it is hypothesized that the protocol results in in vivo immunization of host B lymphocytes (primary immunization) followed by a second in vitro stimulation of B lymphocytes (secondary antigen priming stage) that may improve the ability and propensity of recovered clonal B lymphocytes to secrete an antibody

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monoclonal high affinity monoclonal specific for the target antigen.

Third, it has been observed (as shown herein with IL-6 specific B cells) that the B cell selection protocol reproducibly provides IgG-producing enriched B cells that are, on average, highly selective (antigen specific) for the desired target. Lymphocytes are believed

Antigen-enriched B recovered by these methods contain B lymphocytes capable of producing the desired total complement of epitopic specificities as discussed above.

Fourth, it has been observed that B lymphocyte selection protocols, even when used with small antigens, that is, peptides of 100 amino acids or less, eg, 5-50 amino acids long, reproducibly generates a clonal culture of B lymphocytes that secretes a single high-affinity antibody to the small antigen, eg, a peptide. This is surprising since in general it is quite difficult, it costs a lot of work and sometimes it is not even possible to produce high affinity antibodies for small peptides. Accordingly, the invention can be used to produce therapeutic antibodies to the desired peptide targets, eg, viral, bacterial, or autoantigen peptides, and thus allow the production of monoclonal antibodies with

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very specific binding properties or even the production of a cocktail of monoclonal antibodies for different peptide targets, eg different viral strains. This advantage may be especially useful in the context of the production of a therapeutic or prophylactic vaccine having a desired valency, such as a HPV vaccine that induces protective immunity for different HPV strains.

Fifth, the lymphocyte selection protocol

B, particularly when used with rabbit derived B lymphocytes, tends to reproducibly produce antigen-specific antibody sequences that are very similar to endogenous human immunoglobulins (about 90% similar to the amino acid level) and that contain CDRs that are very similar in length to human immunoglobulins and therefore require little or no sequence modification (typically at most only a few CDR residues can be modified in the main antibody sequence and are not introduced flanking exogenous residues) in order to eliminate potential potential immunogenicity problems. In particular, preferably the recombinant antibody will contain only the host (rabbit) CDR1 and CDR2 residues required for antigen recognition and all of CDR3. Thus, the high antigen-binding affinity of the recovered antibody sequences produced according to the B-lymphocyte and antibody selection protocol remains

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intact or substantially intact even with humanization.

In summary, these methods can be used to produce antibodies that exhibit higher binding affinities with more distinct epitopes by using a more efficient protocol than is already known.

In a specific embodiment, the present invention provides a method of identifying a simple B lymphocyte that secretes an antibody specific for a desired antigen and optionally possesses at least one desired functional property such as affinity, avidity, cytolytic activity, and the like, by a process that includes the following stages:

to. immunizing a host against an antigen;

b. culturing host B lymphocytes;

c. enrich cultured B lymphocytes to increase the frequency of antigen-specific cells;

d. create at least one-celled suspension;

and. culturing a subpopulation from the single-cell suspension under conditions that favor the survival of a single antigen-specific B lymphocyte per culture well;

F. isolating B lymphocytes from the subpopulation; and

g. determine if the simple B lymphocyte produces an antigen-specific antibody.

Typically, these methods will further comprise

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IMPI

MEXICAN INSTITUTE OF "INDUSTRIAL ROPILTY" an additional step of isolation and determination of - II »I I. · * - ~ II sequence, partial or total, of the polypeptide and nucleic acid sequences that encode the desired antibody. These modified sequences or versions or portions thereof can be expressed in desired host cells to produce recombinant antibodies to a desired antigen.

As noted above, the clonal population of B lymphocytes is believed to comprise predominantly B lymphocytes that secrete antibodies that produce an antibody against the desired antigen. It is also believed, based on experimental results obtained with various antigens and with different populations of B lymphocytes, that the B lymphocytes produced by cloning and the isolated antigen-specific B lymphocytes derived therefrom, produced according to the invention, they secrete a monoclonal antibody that typically has high affinity and is furthermore capable of efficiently and reproducibly producing a selection of monoclonal antibodies of greater epitopic variability compared to other methods of obtaining monoclonal antibodies from cultured B lymphocytes specific for antigen. In an exemplary embodiment, the population of immune cells used in said B cell selection methods comes from a rabbit. However, other hosts that produce antibodies, including non-human and human hosts, may alternatively

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D £ LA? RC? I ££> AO MEXICAN INSTITUTE

INDUSTRY!

be used as a source of immune B lymphocytes. It is believed that the use of rabbits as a source of B lymphocytes can improve the diversity of monoclonal antibodies that can be raised by these methods. Also, antibody sequences originating from rabbits according to the invention typically possess sequences with a high degree of sequence identity with human antibody sequences making them favorable for use in humans since they should possess low antigenicity. Over the course of humanization, the final humanized antibody has a much lower foreign / host residue content, typically restricted to a subset of host CDR residues that differ dramatically due to their nature compared to the human target sequence used in grafts . This improves the probability of full recovery of activity in the humanized antibody protein.

Antibody selection methods using an enrichment step described herein include a step for obtaining a cell population containing immune cells from an immunized host. Methods for obtaining a cell population containing immune cells from an immunized host are known in the art and generally include inducing an immune response in a host and culturing cells from the host to obtain one or more cell populations. The

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response can be elicited by immunizing the host against a desired antigen. Alternatively, the host used as the source of such immune cells can naturally be exposed to the desired antigen such as an individual who was infected with a particular pathogen such as a bacterium or virus or alternatively has mounted a cancer-specific antibody response suffered by the individual.

Host animals are well known in the art and include, but are not limited to, guinea pigs, rabbits, mice, rats, non-human primates, humans, as well as other mammals and rodents, chickens, cows, pigs, goats, and sheep.

Preferably the host is a mammal, more preferably a rabbit, mouse, rat, or human. Upon exposure to an antigen, the host produces antibodies as part of the natural immune response to the antigen. As mentioned, the immune response can be of natural origin, as a result of disease, or can be induced by immunization with the antigen. Immunization can be performed by any method known in the art, such as, by one or more injections of the antigen with or without an agent to enhance the immune response, such as a complete or incomplete Freund's adjuvant. In another embodiment, the invention also contemplates intrasplenic immunization. As an alternative to immunizing a host animal in vivo, the method can

398 comprise immunizing an in vitro culture.

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MEXICAN INSTITUTE DS THE INDUSrWAL PROPERTY

,,,, “-'DUPl I-JAL -¾¾. «Host cells o: n

After allowing time for the immune response (eg, as measured by serum antibody detection), animal host cells are cultured to obtain one or more cell populations. In a preferred embodiment, a cultured cell population is analyzed to determine antibody binding strength and / or antibody functionality

A cultured cell population is preferably at least one of spleen, lymph nodes, bone marrow, and / or peripheral blood mononuclear cells (PBMC). Cells can be grown from more than one source and accumulate. Some sources may be preferred for certain antigens. For example, the spleen, lymph nodes, and PBMC are preferred for

IL-6; and lymph nodes are preferred for TNF. The cell population is cultured for about 20 to about 90 days or increments thereof after immunization, preferably for about 50 to about 60 days. A cultured cell population and / or a single-cell suspension thereof can be enriched, analyzed and / or cultured for antibody selection. The frequency of antigen-specific cells within a cultured cell population is normally about 1% to about

5% or increases thereof.

In one embodiment, a single cell suspension of a

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY cultivated cell population is enriched, preferably using Miltenyi beads. From the cultured cell population with an antigen-specific cell frequency of about 1% to about 5%, an enriched cell population is obtained with an antigen-specific cell frequency approaching 100%.

The antibody selection method using an enrichment step includes an antibody production step of at least one antigen-specific cell from an enriched cell population. In vitro antibody production methods are well known in the art and any suitable method can be used. In one embodiment, an enriched cell population, such as an antigen-specific single cell suspension from a cultured cell population, was plated at various cell densities, such as 50, 100, 250, 500, or other increments of 1 to 1000 cells per well. Preferably, the subpopulation comprises no more than about 10,000 cells that secrete antigen-specific antibodies, more preferably about 50-10,000, about 50-5,000, about 501,000, about 50-500, about 50-250 cells that secrete antigen-specific antibodies or increases thereof. These subpopulations are then cultured with a suitable medium (eg, an activated T lymphocyte conditioned medium, in particular 1-5% of a conditioned medium of

400

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY activated rabbit T lymphocytes) in a feeding layer, preferably under conditions that favor the survival of a single proliferating cell that secretes antibodies per culture well. The feeder layer, which generally comprises irradiated cellular material, eg, EL4B cells, does not constitute part of the cell population. Cells are cultured in a suitable medium for a period sufficient for antibody production, for example from about 1 day to about 2 weeks, from about 1 day to about 10 days, at least about 3 days, about 3 to around 5 days, around 5 days to around 7 days, at least around 7 days or increments thereof. In one embodiment, more than one subpopulation is grown simultaneously. Preferably, a single cell that produces antibodies and progeny thereof survives in each well, thereby providing a clonal population of antigen-specific B lymphocytes in each well. At this stage, the immunoglobulin G (IgG) produced by the clonal population is highly correlated with the specificity of the antigen. In a preferred embodiment, IgGs exhibit a correlation with antigen specificity that is greater than about 50%, more preferably greater than 70%, 85%, 90%, 95%, 99%, or increases thereof. See Figure 3 which shows an example of correlation for IL-6. Correlations were shown by placing B lymphocyte cultures under limiting conditions to

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establish simple antigen-specific antibody products per well. Antigen-specific syntheses were compared to general IgG syntheses. Three populations were observed: an IgG that recognized a simple antigen format (biotinylated and direct coating), detectable IgG and antigen recognition without taking into account the immobilization and production of IgG only. The production of

IgG was highly correlated with antigen specificity.

Optionally, a supernatant is collected containing antibodies that can be enriched, analyzed and / or cultivated for the selection of antibodies according to the steps described above. In one embodiment, the supernatant is enriched (preferably by means of an antigen specificity assay, especially an ELISA assay) and / or analyzed to determine the functionality of the antibody.

In another embodiment, the enriched, preferably clonal, population of antigen-specific B lymphocytes from which an above-described supernatant is optionally assayed for the presence of the desired secreted monoclonal antibody is used for the isolation of a few B lymphocytes, preferably a single B lymphocyte which is then tested in an appropriate assay to confirm the presence of a single B lymphocyte that produces antibodies in the clonal population of B lymphocytes.

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modality, about 1 to about 20 cells are isolated from the clonal population of B lymphocytes, preferably less than about 15, 12, 10, 5, or 3 cells, or increments thereof, more preferably a single cell . The assay is preferably carried out by an antigen specificity assay, especially a halogen assay. The halogen assay can be performed with the full-length protein or a fragment thereof. The antibody-containing supernatant can also be analyzed to determine at least one of: antigen binding affinity; agonism or antagonism of ligand binding to antigen, induction or inhibition of proliferation of a specific target cell type; induction or inhibition of lysis of a target cell and induction or inhibition of a biological pathway involving the antigen.

The identified antigen-specific cell can be used to obtain the corresponding nucleic acid sequences encoding the desired monoclonal antibody. (An AluI digestion can confirm that only a single type of monoclonal antibody is produced per well.) As mentioned above, these sequences can be mutated, such as by humanization, to make them suitable for use in human medications.

As mentioned above, the enriched B lymphocyte population used in the process can also

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further enrich, analyze and / or cultivate for the selection of antibodies according to the steps described above which can be repeated or performed in different order. In a preferred embodiment, at least one cell from an enriched, preferably clonal, antigen-specific cell population is isolated, cultured, and used for antibody selection.

Therefore, in one embodiment, the present invention provides a method comprising:

to. culturing a cell population from an immunized host to obtain a cultured cell population,

b. creating at least one single-cell suspension from a cultured cell population;

c. enriching at least one single-cell suspension, preferably by chromatography, to form a first enriched cell population;

d. enriching the first enriched cell population, preferably by the ELISA assay, to form a second enriched cell population that is preferably clonal, that is, contains only a single type of antigen-specific B lymphocyte.

and. enrich the second enriched cell population, preferably by the halogen assay, to form a

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third enriched cell population containing one or few B lymphocytes that produce an antibody specific for a desired antigen; and

F. selecting an antibody produced by an antigen-specific cell isolated from the enriched third cell population.

The method may additionally include one or more steps of analysis of the cultured cell population for the binding strength of the antibody (affinity, avidity) and / or the functionality of the antibody. Suitable assay steps include but are not limited to assay methods that detect: if the antibody produced by the identified antigen-specific B lymphocyte produces an antibody having minimal antigen-binding affinity, if the antibody agonizes or antagonizes binding from a desired antigen to the ligand; whether the antibody induces or inhibits the proliferation of a specific cell type;

if the antibody induces or causes a cytolytic reaction against target cells; if the antibody binds to a specific epitope; and if the antibody modulates (inhibits or agonizes) the specific biological pathway (s) involving the antigen.

Similarly, the method may additionally include one or more steps of analysis of the second enriched cell population to determine antibody binding strength and / or antibody functionality.

The method may additionally include a step of

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sequence polypeptide or He sequences. the corresponding nucleic acid sequence of the selected antibody. The method may also include a step of producing a recombinant antibody using the sequence, a fragment thereof, or a genetically modified version of the selected antibody. Methods for mutating antibody sequences to retain desired properties are well known to those skilled in the art and include humanization, chimerization, single-chain antibody production; these mutation methods can produce recombinant antibodies that possess the desired effector function, immunogenicity, stability, elimination or addition of glycosylation, and the like. The recombinant antibody can be produced by any suitable recombinant cell that includes but is not limited to mammalian cells such as

CHO, COS, BHK, HEK-293, bacterial cells, yeast cells, plant cells, insect cells, and amphibian cells. In one embodiment, the antibodies are expressed in yeast polyploid cells, that is, yeast diploid cells, in particular Pichia.

In one embodiment, the method comprises:

to. immunizing a host against an antigen to produce host antibodies;

b. analyze host antibodies to determine antigen specificity and neutralization;

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IMPI MEXICAN INSTITUTE

b. culturing host B lymphocytes;

d. enriching cultured B lymphocytes to create an enriched cell population with an increased frequency of antigen-specific cells;

and. culturing one or more subpopulations of the enriched cell population under conditions that favor the survival of a single B lymphocyte to produce a clonal population in at least one culture well;

F. determine whether the clonal population produces an antigen-specific antibody.

g. isolate a simple B lymphocyte; and

h. sequencing the nucleic acid sequence of the antibody produced by the simple B lymphocyte.

Antibody humanization methods

In another embodiment of the invention, a method is provided for the humanization of antibody heavy and light chains. In this embodiment, the following method is followed for the humanization of heavy and light chains:

Light chain

one. Identify the amino acid that is the first to follow the signal peptide sequence. This is the beginning of Flanking Region 1. The signal peptide begins at the first initiating methionine and is typically, but not necessarily, 22 amino acids long

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for rabbit light chain protein sequences. The start of the mature polypeptide can also be determined experimentally by sequencing the N-terminal protein or can be predicted using a prediction algorithm. This is also the beginning of Flanking Region 1 as it is usually defined by experts in the field.

Example: Amino acid residue RbtVL 1 in Figure 2, beginning with AYDM.

2. Identify the end of Flanking Region 3. This typically has 86-90 amino acids that follow the beginning of Flanking Region 1 and is typically a cysteine residue preceded by two tyrosine residues. This is the end of Flanking Region 3 as it is usually defined by experts in the field.

Example: Amino acid residue RbtVL 88 in Figure 2, ending with TYYC.

3. Use the rabbit light chain sequence of the polypeptide starting from the beginning of Flanking Region 1 to the end of Flanking Region 3 as defined above and perform a sequence homology search for the most similar antibody protein sequences. This will typically be a search against human germ sequences prior to antibody maturation to reduce the possibility of immunogenicity; however, any human sequence can be used. Typically

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IMPI

INSTITUTO MEXICANO DE LA PROPIEDAD industrial a BLAST-style program can be used to search a sequence database for the most homologous. Human antibody sequence databases can be found in various sources such as NCBI (National Center for

Biotechnological Information).

Example: The RbtVL amino acid sequence of residues numbered 1 to 88 in Figure 2 is compared in BLAST with a germ line database of human antibodies. The first three unique sequences returned are shown in Figure 2 as L12A, VI, and Vx02.

Four. In general, the most homologous human germline variable light chain sequence is then used as the basis for humanization. However, those skilled in the art may decide to use another sequence that is not of higher homology as determined by the homology algorithm, based on other factors including sequence spaces and similarities in the flanking region.

Example: In Figure 2, L12A was the most homologous human germline variable light chain sequence and is used as the basis for humanization of RbtVL.

5. Determine the Flanking Region and CDR array (FRl, FR2, FR3, CDR1 and CDR2) for the human homolog used for light chain humanization. This is using the traditional distribution as described in the subject. Align the rabbit variable light chain sequence with the homolog

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY, while maintaining the distribution of the flanking regions and CDR.

Example: In Figure 2, the RbtVL sequence is aligned with the human homologous sequence L12A and the flanking and CDR domains are indicated.

6. Replace the CDR1 and CDR2 regions of the human homologous light chain sequence with the CDR1 and CDR2 sequences of the rabbit sequence. If there were length differences between the human and rabbit CDR sequences then use all of the rabbit CDR sequences and their lengths. It is possible that the specificity, affinity and / or immunogenicity of the resulting humanized antibody may not be altered if major or minor sequence exchanges are performed or if specific residues are altered, however exchanges as described have been used successfully but are not excluded. the possibility of other changes being allowed.

Example: In Figure 2, the amino acid residues of

CDR1 and CDR2 of the human homologous variable light chain L12A are replaced with the amino acid sequences of CDR1 and CDR2 of the rabbit antibody light chain sequence RbtVL. Flanking regions 1, 2 and 3 of human L12A are unchanged. The resulting humanized sequence is shown below as VLh of residues numbered 1 to 88.

Note that only residues that are different from the

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INS. ϊ. ν7C MFX13.ANO V; - <rcyr ^ TV / ^^. ,, _ "'INDUSTRIAL SiiSX-U · ^, human sequence L12A and therefore are the amino acid residues that come from rabbit. In this example όοΙΊόΠΓΈβ residues are different from the human sequence.

7. Following flanking region 3 of the new hybrid sequence created in Step 6, attach the entire CDR3 of the rabbit light chain antibody sequence. The CDR3 sequence can be of various lengths but is typically 9 to 15 amino acid residues in length. The CDR3 region and the beginning of the next region of flanking region 4 are defined classically and so that they can be identified by those skilled in the art. Typically the beginning of flanking region 4 and therefore after the end of CDR3 consists of the FGGG ... sequence, however there may be some variation in these residues.

Example: In Figure 2, the CDR3 of RbtVL (amino acid residues numbered 8 9 to 100) is added to the end of flanking region 3 in the humanized sequence indicated as

VLh.

8. Flanking Region 4 of the rabbit light chain which is typically the last 11 amino acid residues of the variable light chain and begins as indicated in Step 7 above and typically ends with the amino acid sequence ... VVKR, is replaced with the closest human light chain Flanking Region 4 homolog, usually of the germ sequence. Frequently this region

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IMPI

INSTITUTO MEXICANO D £ LA PRCITEDAD INDUSTRIAL 4 flanking human light chain is of the sequence 'FGGGTKVEIKR'. It is possible that other sequences of human light chain Flanking Region 4 that are not the most homologous or otherwise different, may be used without affecting the specificity, affinity and / or immunogenicity of the resulting humanized antibody. This sequence of the Region

Human light chain flanking 4 is added to the end of the humanized variable light chain sequence that immediately follows the CDR3 sequence from Step 7 above.

This is the end of the variable light chain humanized amino acid sequence.

Example: In Figure 2, flanking region 4 (FR4) of the rabbit light chain sequence RbtVL is shown on a homologous human FR4 sequence. The human FR4 sequence is added to the humanized variable light chain (VLh) sequence just after the end of the CD3 region added in Step 7 above.

Heavy chain

one. Identify the amino acid that is the first to follow the signal peptide sequence. This is the beginning of Flanking Region 1. The signal peptide begins at the first initiation methionine and is typically 19 amino acids in length for rabbit heavy chain protein sequences. Typically, but not

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I

IMST'T 'l'. fj f ^ lC ^ NOT necessarily always, the last 3 amiriciazide residues of a conTJo ^ 'heavy chain signal peptide are ... VQC followed by the start of Flanking Region 1. The start of the mature polypeptide can also be determined experimentally by sequencing the N-terminal protein or it can be predicted using a prediction algorithm. This is also the beginning of Flanking Region 1 as it is usually defined by experts in the field.

Example: Amino acid residue RbtVH 1 in Figure 2, starting with QEQL ...

2. Identify the end of Flanking Region 3. This typically has 95-100 amino acids that follow the beginning of Flanking Region 1 and typically has the ending sequence.

... CAR (although alanine can also be a valine).

This is the end of Flanking Region 3 as it is usually defined by experts in the field.

Example: Amino acid residue RbtVH 98 in Figure 2, ending with ... FCVR.

3. Use the rabbit heavy chain sequence of the polypeptide starting from the beginning of Flanking Region 1 to the end of Flanking Region 3 as defined above and perform a sequence homology search for the most similar antibody protein sequences. This will typically be against a human germ sequence database prior to

413

antibody maturation to reduce the possibility of immunogenicity; however any human sequence can be used. Typically you can use a program like

BLAST to search a sequence database for the most homologous. Human antibody sequence databases can be found at various sources such as NCBI (National Center for Biotechnology Information).

Example: The RbtVH amino acid sequence of residues numbered 1 to 98 in Figure 2 is compared in BLAST with a database of human antibody germ lines. The first three unique sequences returned are shown in Figure 2 as 3-64-04, 3-66-04, and 3-53-02.

Four. In general, the most homologous human germline variable heavy chain sequence is then used as the basis for humanization. However, those skilled in the art may decide to use a sequence other than the most homologous as determined by the homology algorithm, based on other factors including sequence spaces and flanking region similarities.

Example: 3-64-04 in Figure 2, was the most homologous human germline variable heavy chain sequence and is used as the basis for the humanization of RbtVH.

5. Determine the flanking region and CDR array (FR1, FR2, FR3, CDR1, and CDR2) for the human homolog used for heavy chain humanization. This is using the distribution

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MEXICAN INSTITUTE

OF THE PROPERTY

INDUSTRIAL traditional as described in the matter. Align the rabbit variable heavy chain sequence with the human homolog, while maintaining the distribution of the flanking regions and CDR.

Example: In Figure 2, the RbtVH sequence is aligned with the human homologous sequence 3-64-04 and flanking and CDR domains are indicated.

6. Replace the CDR1 and CDR2 regions of the human homologous heavy chain sequence with the CDR1 and CDR2 sequences of the rabbit sequence. If there were length differences between the human and rabbit CDR sequences then use all of the rabbit CDR sequences and their lengths. Additionally, it may be necessary to replace the last three amino acids of human heavy chain flanking region 1 with the last three amino acids of rabbit heavy chain flanking region 1. Typically, but not always, in the rabbit heavy chain flanking region 1 these three residues follow a glycine residue preceded by a serine residue. Furthermore, it may be necessary to replace the final amino acid of human heavy chain flanking region 2 with the final amino acid of rabbit heavy chain flanking region 2. Typically, but not necessarily always, this is a glycine residue preceded by an isoleucine residue in the rabbit heavy chain flanking region 2. It is possible that the specificity, the

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MEXICAN INSTITUTE

FROM THE OWN PROPERTY

INDUSTRIAL antibody

Affinity and / or immunogenicity of the resultant may not be modified if smaller or larger sequence exchanges are performed or if specific residues are modified; however the exchanges as described were used successfully but do not exclude the possibility that other changes are allowed. For example, an amino acid residue of tryptophan typically occurs four residues before the end of the region

Rabbit heavy chain CDR2, whereas human heavy chain CDR2 this residue is typically a serine residue. Changing this rabbit tryptophan residue to a human serine residue in this position has been shown to have little or no effect on the specificity or affinity of the humanized antibody, and thus further minimizes the content of amino acid residues that come from the rabbit sequence in the humanized sequence.

Example: In Figure 2, the amino acid residues of

CDR1 and CDR2 of the human homologous variable heavy chain are replaced by the amino acid sequences of CDR1 and CDR2 of the rabbit antibody light chain sequence RbtVH, except for the sequenced residue which is tryptophan in the rabbit sequence (position number 63 ) and serine at the same position in the human sequence and remains as the human serine residue. In addition to the CDR1 and CDR2 changes, the

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The last three amino acids from flanking region 1 (positions 28-30) as well as the final residue from flanking region 2 (positions 49) are retained as rabbit rather than human amino acid residues. The resulting humanized sequence is shown below as VHh of residues numbered 1 to

98. Note that only residues that are different from the human 3-64-04 sequence are underlined and therefore are amino acid residues that come from rabbit. In this example only 15 of the 98 residues are different from the human sequence.

7. Following flanking region 3 of the new hybrid sequence created in Step 6, attach the entire CDR3 of the rabbit heavy chain antibody sequence. The CDR3 sequence can be of various lengths but is typically 5 to 19 amino acid residues in length. The CDR3 region and the beginning of the next flanking region 4 are defined classically and so that they can be identified by those skilled in the art. Typically the beginning of flanking region 4 and therefore after the end of CDR3 consists of the sequence WGXG ... (where X is normally Q or P); however there may be some variation in these residues.

Example: CDR3 from RbtVH (amino acid residues numbered 99 to 110) is added to the end of flanking region 3 in the humanized sequence indicated as VHh.

8. Flanking region 4 of the rabbit heavy chain

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D £ LA RkC'i'iE? \ D

INDUSTRIAL which are typically the last 11 amino acid residues of

VariableηηίΜβΤηΚΜΜΒΙΙ the variable heavy chain and begins as indicated in Step 7 above and typically ends with the amino acid sequence ... TVSS is replaced with the closest human heavy chain flanking region 4 homolog, usually from the germline sequence. Frequently, this human heavy chain 4 flanking region is of the sequence 'WGQGTLVTVSS. It is possible that other sequences of the human heavy chain flanking region 4 that are not the most homologous or otherwise different, can be used without affecting the specificity, affinity and / or immunogenicity of the resulting humanized antibody. This sequence of human heavy chain flanking region 4 is added to the end of the humanized variable heavy chain sequence that immediately follows the CDR3 sequence from Step 7 above.

This is the end of the variable heavy chain humanized amino acid sequence.

Example: In Figure 2, flanking region 4 (FR4) of the RbtVH rabbit heavy chain sequence is shown on a homologous human heavy FR4 sequence. The human FR4 sequence is added to the humanized variable heavy chain (VHh) sequence just after the end of the CD3 region that is added in Step 7 above.

Methods for the production of antibodies and fragments of

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themselves

The invention is also directed to the production of antibodies described herein or fragments thereof. Recombinant polypeptides corresponding to the antibodies described herein or fragments thereof are secreted from polyploid strains, preferably diploid or tetraploid yeast capable of mating. In one exemplary embodiment, the invention is directed to methods for the production of these recombinant polypeptides in secreted form for prolonged periods using cultures comprising poiiploid yeast, that is, at least several days to a week, more preferably at least a month or several months and even more preferably at least 6 months to a year or more. These poiiploid yeast cultures will express at least 10-25 mg / liter of the polypeptide, more preferably at least 50-250 mg / liter, even more preferably at least 500-1000 mg / liter and more preferably one gram per liter or more than / of the recombinant polypeptide / s.

In one embodiment of the invention, a pair of genetically labeled yeast haploid cells are transformed with expression vectors comprising subunits of a desired heteromultimeric protein. A haploid cell comprises a first expression vector and a second haploid cell comprises a second expression vector. In another embodiment, the diploid yeast cells will be

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transformed with one or more expression vectors that provide the expression and secretion of one or more of the recombinant polypeptides. In yet another embodiment, a single haploid cell can be transformed with one or more vectors and used to produce a polyploid yeast by fusion or pairing strategies. In yet another embodiment, a diploid yeast culture can be transformed with one or more vectors that provide for the expression and secretion of a desired polypeptide / s. These vectors can comprise vectors, eg. Linearized plasmids or other linear DNA products that integrate the yeast cell genome randomly, through homologous recombination or using a recombinase such as Cre / Lox or Flp / Frt. Optionally, additional expression vectors can be introduced into haploid or diploid cells; or the first or second expression vector may comprise additional coding sequences for heterotrimer synthesis; heterotetramers; etc. The expression levels of the non-identical polypeptides can be individually calibrated and adjusted through appropriate selection, vector copy number, promoter strength and / or induction, and the like. Transformed haploid cells genetically cross or fuse. The resulting diploid or tetraploid strains are used to produce and secrete fully assembled and biologically functional proteins, humanized antibodies described in

420

D £ THE INDUSTRIAL PROPERTY present or fragments thereof.

The use of diploid or tetraploid cells for protein production provides unexpected benefits.

Cells can be cultured for production purposes, that is, scaled up, and for prolonged periods of time, under conditions that can be detrimental to haploid cell culture, whose conditions may include high cell density; culture in minimal media; cultivation at low temperatures; Stable culture in the absence of selective pressure and which can provide maintenance of heterologous gene sequence integrity and maintenance of a high level of expression over time. Without wishing to limit themselves to this, the inventors theorize that these benefits may arise, at least in part, from the creation of diploid strains from two different parental haploid strains. Such haploid strains can comprise several minor autotrophic mutations; These mutations complement each other in the diploid or tetraploid, enabling growth and improved production under highly selective conditions.

Mating capable haploid yeast cells provide a genetic method that allows mating of subunits of a desired protein. The haploid yeast strains are transformed with each of the two expression vectors, a first vector to direct the synthesis of a polypeptide chain and a second vector to

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INDUSTRIAL direct the synthesis of a second chain of non-identical polypeptides. The two haploid strains pair to provide a diploid host where optimized target protein production can be obtained.

Optionally, non-identical coding sequences / s are provided. Such sequences may be present in additional expression vectors or in the first or second expression vector. As is known in the art, multiple coding sequences can be independently expressed from individual promoters; or they can be expressed in a coordinated manner through the inclusion of an internal ribosome entry site or IRES which is an element that promotes direct internal ribosome entry to the initiation codon such as ATG of a cistron (a region encoding a protein) causing an independent gene translation of cap. Functional IRES elements in yeast are described in Thompson et al. (2001) PNAS 98: 12866-12868.

In one embodiment of the invention, the antibody sequences are produced in combination with a J secretory chain, which provides improved IgA stability (see US Patent Nos. 5,959,177; and 5,202,422).

In a preferred embodiment, the two haploid yeast strains are each auxotrophic and require a complement of media for haploid cell culture. The pair of auxotrophs are complementary, so that the product

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Diploid MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY will grow in the absence of the complements required for haploid cells. Many of these genetic markers are known in yeast, and include requirements for amino acids (eg, met, lys, his, arg, etc.), nucleosides (eg, ura3, adel, etc.); and the like. Amino acid markers will be preferred for the methods of the invention. Alternatively, diploid cells containing the desired vectors can be selected by other means, eg, by the use of other markers such as fluorescent green protein, antibiotic resistant genes, various dominant selectable markers, and the like.

Two transformed haploid cells can be genetically crossed and diploid strains arising from this mating event can be selected for their hybrid nutritional requirements and / or antibiotic resistance spectrum. Alternatively, populations of the two transformed haploid strains undergo spheroplast and fuse and regenerate and select diploid progeny. By any method, diploid strains can be identified and selectively grown based on their ability to grow in media other than their parents. For example, diploid cells can be grown in minimal medium that can include antibiotics. The diploid synthesis strategy has certain advantages. Diploid strains have the potential to produce improved levels of protein

423

heterologous through broader complementation for ____ underlying mutations that may impact the production and / or secretion of recombinant protein. Furthermore, once stable strains are obtained, any antibiotics used to select those strains need not necessarily be continuously present in the culture medium.

As noted above, in some embodiments, a haploid yeast can be transformed with one or multiple vectors and paired or fused with an untransformed cell to produce a diploid cell containing the vector or vectors. In other embodiments, a yeast diploid cell can be transformed with one or more vectors that provide expression and secretion of a desired heterologous polypeptide via the yeast diploid cell.

In one embodiment of the invention, two haploid strains are transformed with a library of polypeptides, eg, a library of heavy or light chain antibodies. Transformed haploid cells that synthesize the polypeptides mate with complementary haploid cells. The resulting diploid cells are analyzed to determine the functional protein. Diploid cells provide a means of quickly, conveniently and inexpensively assembling a large number of combinations of polypeptides for functional testing. This technology is especially applicable for the creation of heterodimeric protein products, where

424

IMPIC.,

MEXICAN INSTITUTE »Z * '' Rr'p'táJ DE ΙΑ PROPIEDAD -. T-tijiJ '

INDUSTRIAL optimized subunit synthesis levels are critical for the expression and secretion of functional proteins.

In another embodiment of the invention, the ratio of the expression level of the two subunits is regulated in order to maximize the creation of products. Heterodimeric subunit protein levels have previously been shown to impact the creation of end products (Simmons LC, J Immunol Methods. May 2002 1; 263 (1-2): 13347). Regulation can be accomplished prior to the mating step by selecting a marker present in the expression vector. The level of expression can be increased by stably increasing the copy number of the vector. In some cases, it may be desired to increase the level of one chain relative to another so as to achieve a balanced ratio between the subunits of the polypeptide. Antibiotic resistance markers are useful for this purpose, eg.

Zeocin ™ resistance marker (phleomycin), resistance to

G418, etc., and provide an enrichment medium for strains containing multiple integrated copies of an expression vector in a strain by selecting transformers resistant to higher levels of Zeocin ™ (phleomycin) or G418. The right relationship, eg. 1: 1, 1: 2, etc. Subunit genes may be important for efficient protein production. Even when the same promoter is used to transcribe both subunits, many other factors contribute to the level

425

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MEXICAN INSTITUTE i-—, -i OF THE PROPERTY UiE-T-T '. T · '<^ uXtríir'T -'- Ss ·'1The final protein is expressed and can therefore be useful to increase the number of copies of one encoded gene relative to the other. Alternatively, diploid strains that produce higher levels of a polypeptide relative to single copy vector strains are created by pairing two haploid strains and both have multiple copies of the expression vectors.

Host cells are transformed with the aforementioned expression vectors, paired to form diploid strains and grown in conventionally modified nutrient media as appropriate for promoter induction, selection of transformers, or amplification of genes encoding desired sequences. A number of minimum media suitable for yeast culture are known in the art. Any of these media can be supplemented as needed with salts (such as sodium, calcium, magnesium, and phosphate chloride), buffers (such as phosphate, HEPES), nucleosides (such as adenosine and thymidine), antibiotics, trace elements, and glucose or an equivalent energy source.

Any other necessary supplements may also be included in appropriate concentrations known to those of skill in the art. The culture conditions, such as temperature, pH and the like are those previously used with the host cell selected for expression and will be apparent to those skilled in the art.

The secreted proteins are recovered from the

426

culture. A protease inhibitor, such as phenyl methyl sulfonyl fluoride (PMSF) can be useful to inhibit proteolytic degradation during purification and antibiotics can be included to prevent the cultivation of accidental contaminants. The composition can be concentrated, filtered, dialyzed, etc., using methods known in the art.

The diploid cells of the invention are grown for production purposes. Such production purposes desirably include cultivation in minimal media lacking preformed amino acids and other complex biomolecules, eg, media comprising ammonia as a source of nitrogen and glucose as a source of energy and carbon, and salts as a source of phosphate, calcium and the like. Preferably, such means of production lack selective agents such as antibiotics, amino acids, purines, pyrimidines, etc. Diploid cells can be grown at high cell density, for example at least about 50 g / L; more usually at least around 100 g / L; and at least about 300, about 400, about 500 g / L or more.

In one embodiment of the invention, the cells in question are cultured for production purposes at low temperatures that can be lowered during the logarithmic phase, during the stationary phase, or both. The term low temperature refers to temperatures of at least around 15 ° C, more usually at least around 17 ° C

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

and they can be around 20 ° C and normally not higher than around 25 ° C, more usually not higher than around 22 ° C. In another embodiment of the invention, the low temperature is usually not greater than about 28 ° C. The culture temperature can impact the production of full length secreted proteins in production cultures and reducing the culture growth temperatures can strongly increase the yield of the intact product. The decreased temperature appears to assist intracellular traffic through the folding and post-translational processing pathways used by the host to generate the target product along with reduced degradation of cellular protease.

The methods of the invention provide the expression of secreted protein preferably activates a mammalian protein. In one embodiment, the secreted active antibodies, as used herein, refer to a correctly folded multimer of at least two appropriately arranged strands in pairs and accurately binding to their cognate antigen. Expression levels of active proteins are normally at least about 10-50 mg / liter of the culture, more usually at least about 100 mg / liter, preferably at least about 500 mg / liter and can be 1000 mg / liter or more.

The methods of the invention can provide a

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY increased stability of host and heterologous coding sequences during production. Stability is demonstrated, for example, by maintaining high levels of time expression, where the starting expression level decreases by no more than about 20%, typically not more than about 10%, and may decrease by no more 5% more than about 20 duplications, 50 duplications, 100 duplications or more.

The stability of the strains also provides maintenance of the integrity of heterologous gene sequences over time, where the sequence of the active coding sequence and the necessary elements of transcriptional regulation are maintained in at least around 99% of diploid cells, normally in at least about 99.9% of diploid cells and preferably in at least about 99.99% of diploid cells more than about 20 duplicates, 50 duplications, 100 duplications or more. Preferably, substantially all diploid cells maintain the active coding sequence sequence and the necessary elements of transcriptional regulation.

Other methods for the production of antibodies are well known to those skilled in the art. For example, methods for the production of chimeric antibodies are now well known in the art (See, eg, Patent

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I

MEXICAN INSTITUTE OF PROPERTY

US INDUSTRIAL No. 4,816,567 to Cabilly et al .; Morrison et al.,

PNAS USA, 81: 8651-55 (1984); Neuberger, MS et al.,

Nature, 314: 268-270 (1985); Boulianne, GL et al., Nature,

312: 643-46 (1984), the disclosures of which are incorporated herein by reference in their entirety).

Also, other methods for the production of humanized antibodies are now well known in the art (See, eg, US Patent No. 5,530,101, 5,585,089,

5,693,762, and 6,180,370 to Queen et al; US patent

Nos. 5,225,539 and 6,548,640 to Winter; US Patent Nos. 6,054,297, 6,407,213 and 6,639,055 to Carter et al; Adair US Patent No. 6,632,927; Jones, PT et al,

Nature, 321: 522-525 (1986); Reichmann, L., et al, Nature, 332: 323-327 (1988); Verhoeyen, M, et al, Science, 239: 1534-36 (1988), the disclosures of which are incorporated herein by reference in their entirety).

Polypeptides of the antibody of the invention possessing IL-6 binding specificity can also be produced by constructing, using standard techniques well known to those skilled in the art, an expression vector containing an operon and a DNA sequence that encodes an antibody heavy chain where the DNA sequence encoding the CDRs necessary for the specificity of the antibody comes from a non-human cellular source, preferably a source of rabbit B lymphocytes, while

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IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY that the DNA sequence that encodes the remaining parts of the antibody chain comes from a human cellular source.

A second expression vector is produced using the same conventional means well known to those skilled in the art, said expression vector contains an operon and a DNA sequence encoding an antibody light chain where the DNA sequence encoding the necessary CDRs for the specificity of the antibody it comes from a non-human cellular source, preferably a rabbit B lymphocyte source, while the DNA sequence encoding the remaining parts of the antibody chain comes from a human cellular source

Expression vectors are transfected into a host cell by standard techniques well known to those skilled in the art to produce a transfected host cell; this transfected host cell cultured by standard techniques well known to those skilled in the art to produce such antibody polypeptides.

The host cell can be transected in conjunction with the two expression vectors described above, the first expression vector contains DNA encoding an operon and a light chain-derived polypeptide, and the second vector contains DNA encoding an operon and a derived polypeptide. from a heavy chain. The two vectors contain

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MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

select different markers, but preferably achieve substantially equal expression of heavy and light chain polypeptides. Alternatively, a simple vector can be used, this vector includes DNA encoding both the heavy and light chain polypeptides. Coding sequences for heavy and light chains may comprise cDNA.

The host cells used to express the antibody polypeptides can be bacterial cells, such as E.coli, or a eukaryotic cell. In a particularly preferred embodiment of the invention, a mammalian cell of a well-defined type can be used for this purpose, such as a myeloma cell or Chinese hamster ovary (CHO) cell line.

General methods by which vectors can be constructed, transfection methods required to produce the host cell, and culture methods required to produce the antibody polypeptides from said host cells, all include conventional techniques. Although preferably the cell line used to produce the antibody is a mammalian cell line, any other suitable cell line, such as a bacterial cell line such as a bacterial strain originating from E.coli, or a yeast cell line, can used alternatively.

Similarly, once the polypeptides of

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IΜ ΡI

MEXICAN INSTITUTE OF THE INDUSTRIAL PRCPICCAD

Antibodies produced can be purified according to standard procedures of the art, such as for example, cross flow filtration, ammonium sulfate precipitation, column affinity chromatography, and the like.

The antibody polypeptides described herein can also be used for the design and synthesis of peptide or non-peptide mimetics that may be useful for the same therapeutic applications as the antibody polypeptides of the invention. See, for example, Saragobi et al, Science, 253: 792-795 (1991), the contents of which are incorporated by reference in their entirety.

Analysis tests

The invention also includes assays for assays designed to assist in the identification of IL-6 associated diseases and disorders in patients displaying symptoms of an IL-6 associated disease or disorder.

In one embodiment of the invention, the anti-IL-6 antibodies of the invention or IL-6 binding fragments thereof, are used to detect the presence of IL-6 in a biological sample obtained from a patient exhibiting symptoms of a IL-6 associated disease or disorder. The presence of IL-6 or high levels of it when compared to pre-disease levels of IL-6 in a comparable biological sample, may be beneficial for the diagnosis of a

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IL-6 associated disease or disorder.

Another embodiment of the invention provides a diagnostic test or assay to assist in the diagnosis of IL-6 associated diseases or disorders in patients exhibiting symptoms of an IL-6 associated disease or disorder identified herein, comprising testing the level expression of IL-6 in a biological sample from said patient using a post-translationally modified anti-IL-6 antibody or a binding fragment thereof. The anti-IL-6 antibody or the binding fragment thereof may be post-translationally modified to include a detectable moiety as previously stated in the description.

The level of IL-6 in the biological sample is determined using a modified anti-IL-6 antibody or a binding fragment thereof as set forth herein and comparing the level of IL-6 in the biological sample against a level IL-6 standard (eg, the level in normal biological samples). The skilled physician will understand that there may be some variability between normal biological samples and will take this into account when evaluating the results.

The aforementioned assay may also be useful in monitoring a disease or disorder, where the level of IL-6 obtained in a biological sample from a patient believed to have an IL-6 associated disease or disorder is compared to the level IL-6 in previous biological samples

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IMPI

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL in the same patient, in order to establish whether the level of IL-6 in that patient has changed, for example, with a treatment regimen.

The invention is also directed to an in vivo imaging method that detects the presence of IL-6 expressing cells comprising administering a diagnostic effective amount of a diagnostic composition. Such in vivo imaging is useful for imaging and diagnosing tumors and metastases that express IL-6 and inflammatory sites that express IL-6, for example, and can be used as part of a planning regimen to design a protocol. effective treatment for arthritis or cancer. The treatment protocol may include, for example, one or more radiation, chemotherapy, cytokine therapy, gene therapy, and antibody therapy, as well as an anti-IL-6 antibody or fragment thereof.

A skilled physician will understand that a biological sample includes, but is not limited to, serum, plasma, urine, saliva, mucosa, pleural fluid, synovial fluid, and spinal fluid.

Methods for improving or reducing the symptoms of, or treating, or preventing diseases and disorders associated with, IL-6.

In one embodiment of the invention, the anti-IL-6 antibodies described herein, or fragments thereof, are

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VI Ρ

UX A MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL 'useful for improving or reducing symptoms of, or treating or preventing diseases and disorders associated with IL-6. The anti-IL-6 antibodies described herein, or fragments thereof, may also be administered in a therapeutically effective amount to patients in need of treatment for IL-6 associated diseases and disorders in the form of a pharmaceutical composition such as It is described in more detail below.

In one embodiment of the invention, the IL6 antagonists described herein are useful for improving or reducing symptoms of, or treating or preventing diseases and disorders associated with elevated C-reactive protein (CRP). Such diseases include any disease presenting chronic inflammation, eg, rheumatoid arthritis, juvenile rheumatoid arthritis, psoriasis, psoriatic arthropathy, ankylosing spondylitis, lupus erythematosus, Crohn's disease, ulcerative colitis, dermatomyositis, polymyositis, polymyalgia rheumatica, giant cell arteritis, vasculitis, polyarteritis nodosa, Wegener's granulomatosis, Kawasaki disease, isolated vasculitis of the central nervous system, ChurgStrauss arteritis, microscopic polyarteritis, microscopic polyangiitis, Henoch-Schonlein purpura, essential cryoglobulinemic vasculitis, rheumatoid vasculitis, cryoglobulinemia, systemic, pemphigus, recurrent polychondritis, Behcet's disease, arteritis of

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Takayasu, multiple, ischemic heart disease, stroke,

MEXICAN INSTITUTE OF THE P «.O?! AGE η INDUSTRIAL

sepsis, vasculitis caused by viral infections (eg hepatitis B, hepatitis C, HIV, cytomegalovirus, Epstein-Barr virus, Parvo B19 virus, etc.), Buerger's disease, cancer, advanced cancer, osteoarthritis, systemic sclerosis, syndrome CREST, Reiter's disease, bone disease

Paget, Sjogran syndrome, type 1 diabetes, type 2 diabetes, familial Mediterranean fever, autoimmune thrombocytopenia, autoimmune hemolytic anemia, autoimmune thyroid disease, pernicious anemia, vitiligo, alopecia areata, primary biliary cirrhosis, chronic autoimmune active hepatitis, alcoholic cirrhosis, hepatitis virus including hepatitis B and C, other organ-specific autoimmune diseases, burns, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, allergic asthma, other allergic conditions, or any combination thereof. In one embodiment of the invention, the anti-IL-6 antibodies described herein, or fragments thereof, are useful for improving or reducing symptoms of, or preventing diseases and disorders associated with reduced serum albumin, eg. , rheumatoid arthritis, cancer, advanced cancer, liver disease, kidney disease, inflammatory bowel disease, celiac disease, trauma, burns, other diseases associated with reduced serum albumin or any combination thereof.

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IMP

MEXICAN INSTITUTE OF PROPERTY INUUSTIU / J.

In another embodiment of the invention, the anti-IL-6 antibodies described herein, or fragments thereof, are administered to a patient in combination with another active agent. For example, an anti-IL-6 antibody or antibody fragment can be co-administered with one or more chemotherapeutic agents, such as VEGF antagonists, EGFR antagonists, platins, taxols, irinotecan, 5fluorouracil, gemcitabine, leucovorin, spheroids, cyclophosphamide, melphalan, vinca alkaloids (eg. , vinblastine, vincristine, vindesine, and vinorelbine), mustines, tyrosine kinase inhibitors, radiation therapy, sex hormone antagonists, selective androgen receptor modulators, selective estrogen receptor modulators, PDGF antagonists, TNF antagonists, IL1 antagonists, interleukins (eg. IL-12 or IL-2), IL12R antagonists, toxin-conjugated monoclonal antibodies, specific monoclonal antibodies against tumor antigens, Erbitux ™, Avastin ™, Pertuzumab, antiCD20 antibodies, Rituxan®, ocrelizumab, ofatumumab, DXL625, Herceptin®, or any combinations thereof.

In one embodiment of the invention, the anti-IL-6 antibodies described herein, or fragments thereof, are useful for improving or reducing symptoms of, or treating or preventing diseases and disorders associated with fatigue. The diseases and disorders associated with fatigue include

438

INDUSTRIAL PROPERTY but not limited to, general fatigue, exercise-induced fatigue, cancer-related fatigue ^ fibromyalgia, inflammatory disease-related fatigue, and chronic fatigue syndrome. See, for example, Esper DH, et al, The cancer cachexia syndrome: a review of metabolic and clinical manifestations, Nutr Clin Pract., August 2005/20 (4): 369-76;

Vgontzas AN, et al, IL-6 and its circadian secretion in humans, Neuroimmunomodulation, 2005; 12 (3) .Ί31-40; Robson-Ansley, PJ, et al, Acute interleukin-6 administration impairs athletic performance in healthy, trained male runners, Can J Appl Physiol., August 2004; 29 (4): 411-8; Shephard RJ., Cytokine responses to physical activity, with particular reference to IL-6: sources, actions, and clinical implications, Crit Rev Immunol., 2002; 22 (3): 165-82; Arnold, MC, et al, Using an interleukin-6 challenge to evalúate neuropsychological performance in chronic fatigue syndrome, Psychol Med., August 2002; 32 (6): 1075-89; Kurzrock R., The role of cytokines in cancer-related fatigue, Cancer, September 15, 2001; 92 (6 Suppl): 1684-8; Nishimoto N, et al, Improvement in Castleman's disease by humanized anti-interleukin-6 receptor antibody therapy, Blood, January 1, 2000; 95 (1): 56-61; Vgontzas AN, et al, Circadian interleukin-6 secretion and quantity and depth of sleep, J Clin Endocrinol Metab., August 1999; 84 (8): 26037 and Spath-Schwalbe E, et al, Acute effects of recombinant human interleukin 6 on endocrine and central nervous sleep functions

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MEXICAN INSTITUTE OF THE ROWEDAD

INDUSTRIAL

in healthy raen, J Clin Endocrinol Metab., May 1998; 83 (5): 1573-9, the disclosures of which are incorporated herein by reference in their entirety.

In a preferred embodiment of the invention, the anti-IL-6 antibodies described herein, or fragments thereof, are useful for improving or reducing symptoms of, or treating or preventing cachexia. The diseases and disorders associated with cachexia include, but are not limited to, cancerous cachexia, cardiac cachexia, respiratory cachexia, renal cachexia, and age-related cachexia. See, for example, Barton, BE., Interleukin-6 and new strategies for the treatment of cancer, hyperproliferative diseases and paraneoplastic syndromes, Expert Opin Ther Targets, August 2005; 9 (4): 737-52; Zaki MH, et al, CNTO 328, a monoclonal antibody to IL-6, inhibits human tumor-induced cachexia in nude mice, Int J Cancer, September 10, 2004; 111 (4): 592-5, - Trikha M, et al, Targeted anti-interleukin-6 monoclonal antibody therapy for cancer: a review of the rationale and clinical evidence, Clin Cáncer Res., October 15, 2003/9 ( 13).-4653-65; Lelli G, et al, Treatment of the cancer anorexia-cachexia syndrome: a critical reappraisal, J Chemother., June 2003; 15 (3): 220-5, - Argiles JM, et al,

Cytokines in the pathogenesis of cancer cachexia, Curr Opin Clin Nutr Metab Care, July 2003; 6 (4): 401-6, - Barton BE.,

IL-6-like cytokines and cancer cachexia: consequences of

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IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY chronic inflammation, Immunol Res., 2001/23 (1): 41-58; Yamashita

JI, et al, Medroxyprogesterone acétate and cancer cachexia:

interleukin-6 involvement, Breast Cancer, 2000/7 (2): 130-5; Yeh

SS, et al, Geriatric cachexia: the role of cytokines, Am J Clin

Nutr., August 1999; 70 (2): 183-97, - Strassmann G, et al,

Inhibition of experimental cancer cachexia by anti-cytokine and anti-cytokine-receptor therapy, Cytokines Mol Ther., June

nineteen ninety five; l (2): 107-13; Fujita J, et al, Anti-interleukin-6 receptor antibody prevents muscle atrophy in colon-26 adenocarcinomabearing mice with modulation of lysosomal and ATP-ubiquitindependent proteolytic pathways, Int J Cáncer, 27 Nov 1996/68 (5): 637-43 ; Tsujinaka T, et al, Interleukin 6 receptor antibody inhibits muscle atrophy and modulates proteolytic systems in interleukin 6 transgenic mice, J Clin Invest., January 1, 1996/97 (1): 244-9 / Emilie D, et al, Administration of an anti-interleukin-6 monoclonal antibody to patients with acquired immunodeficiency syndrome and lymphoma: effect on lymphoma growth and on B clinical Symptoms, Blood, 1994 Oct

15/84 (8): 2472-9, - and Strassmann G, et al, Evidence for the involvement of interleukin 6 in experimental cancer cachexia, J Clin Invest., 1992 May; 89 (5): 1681-4; the descriptions of which are incorporated herein by reference in their entirety.

In another embodiment of the invention, the anti-IL-6 antibodies described herein, or fragments thereof, are useful for improving or reducing symptoms of, or treating or

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prevent autoimmune diseases and disorders. Diseases and disorders associated with autoimmunity include, but are not limited to, rheumatoid arthritis, systemic lupus erythematosus (SLE), systemic juvenile idiopathic arthritis, psoriasis, psoriatic arthropathy, ankylosing spondylitis, inflammatory bowel disease (IBD), rheumatic polymyalgia, arteritis giant cell disease, autoimmune vasculitis, graft versus host disease (GVHD), Sjogren's syndrome, adult-onset Still's disease. In a preferred embodiment of the invention, the humanized anti-IL-6 antibodies described herein, or fragments thereof, are useful for improving or reducing the symptoms of, or treating or preventing rheumatoid arthritis and systemic juvenile idiopathic arthritis. See for example Nishimoto

N., Clinical studies in patients with Castleman's disease,

Crohn's disease, and rheumatoid arthritis in Japan, Clin Rev Allergy Immunol., 2005 Jun; 28 (3): 221-30; Nishimoto N, et al,

Treatment of rheumatoid arthritis with humanized antiinterleukin-6 receptor antibody: a multicenter, double-blind, placebo-controlled trial, Arthritis Rheum., 2004

Jun; 50 (6): 1761-9; Choy E., Interleukin 6 receptor as a target for the treatment of rheumatoid arthritis, Ann Rheum Dis., 2003 Nov; 62 Suppl 2: ii68-9; Nishimoto N, et al, Toxicity, pharmacokinetics, and dose-finding study of repetitive treatment with the humanized anti-interleukin 6 receptor

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INSTITUTO ME / JCANO CE U INDUSTRIAL PROPERTY antibody MRA in rheumatoid arthritis. Phase I / II clinical study, J Rheumatol., 2003 Jul; 30 (7): 1426-35; Mihara M, et al,

Humanized antibody to human interleukin-6 receptor inhibits the development of collagen arthritis in cynomolgus monkeys, Clin

Immunol., 2001 Mar; 98 (3): 319-26; Nishimoto N, et al, Antiinterleukin 6 receptor antibody treatment in rheumatic disease,

Ann Rheum Dis., 2000 Nov; 59 Suppl l: i21-7; Tackey E, et al, Rationale for interleukin-β blockade in systemic lupus erythematosus, Lupus, 2004; 13 (5) :: 339-43; Finck BK, et al,

Interleukin 6 promotes murine lupus in NZB / NZW F1 mice, J Clin

Invest., 1994 Aug; 94 (2): 585-91; Kitani A, et al,

Autostimulatory effects of IL-6 on excessive B cell differentiation in patients with systemic lupus erythematosus: analysis of IL-6 production and IL-6R expression, Clin Exp

Immunol., 1992 Apr; 88 (1): 75-83; Stuart RA, et al, Elevated serum interleukin-6 levels associated with active disease in systemic connective tissue disorders, Clin Exp Rheumatol., 1995 Jan-Feb; 13 (l): 17-22; Mihara M, et al, IL-6 receptor blockage inhibits the onset of autoimmune kidney disease in NZB / W F1 mice, Clin Exp Immunol., 1998 Jun; 12 (3}: 397-402; Woo P, et al, Open label phase II trial of single, ascending doses of MRA in Caucasian children with severe systemic juvenile idiopathic arthritis: proof of principie of the efficacy of IL-6 receptor blockade in this type of arthritis and demonstration of prolonged clinical improvement, Arthritis Res Ther.,

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2005; 7 (6): RI281-8. Electronic publication September 15,

2005; Yokota S, et al, Clinical study of tocilizumab in children with systemic-onset juvenile idiopathic arthritis,

Clin Rev Allergy Immunol., June 2005; 28 (3): 231-8; Yokota S, et al, Therapeutic efficacy of humanized recombinant antiinterleukin-β receptor antibody in children with systemic-onset juvenile idiopathic arthritis, Arthritis Rheum., March 2005; 52 (3): 818-25; de Benedetti F, et al, Targeting the interleukin-β receptor: a new treatment for systemic juvenile idiopathic arthritis ?, Arthritis Rheum., March 2005/52 fd,) .'687-93; De Benedetti F, et al, Is systemic juvenile rheumatoid arthritis an interleukin 6 mediated disease ?, J Rheumatol., February 1998; 25 (2): 203-7; Ishihara K, et al, IL6 in autoimmune disease and chronic inflammatory proliferative disease, Cytokine Growth Factor Rev., August-October 2002; 13 (4-5): 357-68; Gilhar A, et al, In vivo effects of cytokines on psoriatic skin grafted on nude mice: involvement of the tumor necrosis factor (TNF) receptor, Clin Exp Immunol., October

nineteen ninety six; 106 (1): 134-42; Spadaro A, et al, Interleukin-6 and soluble interleukin-2 receptor in psoriatic arthritis: correlations with clinical and laboratory parameters, Clin Exp Rheumatol., July-August 1996; 14 (4): 413-6; Ameglio F, et al,

Interleukin-6 and tumor necrosis factor levels decrease in the suction blister fluids of psoriatic patients during effective therapy, Dermatology, 1994; 189 (4): 359-63; Wendling D, et al,

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Combination therapy of anti-CD4 and anti-IL-6 monoclonal antibodies in a case of severe spondylarthropathy, Br J

Rheumatol., December 1996; 35 (12). * 1330; Gratacos J, et al,

Serum cytokines (IL-6, TNF-alpha, IL-1 beta and IFN-gamma) in ankylosing spondylitis: a cióse correlation between serum IL-6 and disease activity and severity, Br J Rheumatol., October

1994; 33 (10): 927-31; Ito H., Treatment of Crohn's disease with anti-IL-6 receptor antibody, J Gastroenterol., March 2005/40

Suppl 16: 32-4; Ito H, et al, A pilot randomized trial of a human anti-interleukin-6 receptor monoclonal antibody in active Crohn's disease, Gastroenterology, April 2004; 126 (4): 989-96; discussion 947; Ito H., IL-6 and Crohn's disease, Curr Drug Targets Inflamm Allergy, June 2003; 2 (2): 12530; Ito H, et al, Anti-IL-6 monoclonal receptor antibody inhibits leukocyte recruitment and promotes T-cell apoptosis in a murine model of

Crohn's disease, J Gastroenterol., November 2002/37 Suppl

14: 56-61; Ito H., Anti-interleukin-6 therapy for Crohn's disease, Curr Pharm Des., 2003; 9 (4). * 295-305; Salvarani C, et al, Acute-phase reactants and the risk of relapse / recurrence in polymyalgia rheumatica: a prospective follow-up study,

Arthritis Rheum., 2005 Feb 15; 53 (1): 33-8; Roche NE, et al,

Correlation of interleukin-6 production and disease activity in polymyalgia rheumatica and giant cell arteritis, Arthritis

Rheum., September 1993; 36 (9): 1286-94; Gupta M, et al, Cytokine modulation with immune gamma-globulin in peripheral

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blood of normal children and its implications in Kawasaki disease treatment, J Clin Immunol., 2001 May; 21 (3): 193-9; Noris M, et al, Interleukin-6 and RANTES in Takayasu arteritis: a guide for therapeutic decisions ?, Circulation, 1999 Jul

6; 100 (1): 55-60; Besbas N, et al, The role of cytokines in Henoch Schonlein purpura, Scand J Rheumatol., 1997; 26 (6): 45660; Hirohata S, et al, Cerebrospinal fluid interleukin-6 in Progressive Neuro-Behcet's syndrome, Clin Immunol

Immunopathol., January 1997; 82 (1): 12-7; Yamakawa Y, et al, Interleukin-β (IL-6) in patients with Behcet's disease, J Dermatol Sci., March 1996; 11 (3): 189-95; Kim DS., Serum interleukin-6 in Kawasaki disease, Yonsei Med J., June 1992; 33 (2): 183-8; Lange, A., et al, Cytokines, adhesion molecules (E-selectin and VCAM-1) and graft-versus-host disease, Arch. Immunol Ther Exp., 1995, 43 (2): 99-105; Tanaka,

J., et al, Cytokine gene expression after allogeneic bone marrow transplantation, Leuk. Lymphoma, 1995 16 (5-6): 413-418; Dickenson, AM, et al, Predicting outcome in hematological stem cell transplantation, Arch Immunol Ther Exp., 2002 50 (6): 371-8; Zeiser, R, et al, Immunopathogenesis of acute graft-versus-host disease: implications for novel preventive and therapeutic strategies, Ann Hematol., 2004 83 (9): 551-65, - Dickinson, AM, et al, Genetic polymorphisms predicting the outcome of bone marrow transplants, Br. J Haematol., 2004 127 (5): 479-90, - and

Scheinberg MA, et al, Interleukin 6: a possible marker of

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INSTΠ UTO .Μ ΪΧ! CANO D £ LA rEOP.'c'DAD h'-OUL I'IUAL disease activity in adult onset Still's disease, Clin Exp Rheumatol., November-December 1996, -14 (6): 653-5, the descriptions of which are incorporated in its entirety herein for reference.

In another embodiment of the invention, the anti-IL-6 antibodies described herein, or fragments thereof, are useful for improving or reducing symptoms of, or treating or preventing diseases and disorders associated with the bone system. Diseases and disorders associated with the bone system include, but are not limited to, osteoarthritis, osteoporosis, and Paget's disease of bone. In a preferred embodiment of the invention, the humanized anti-IL-6 antibodies described herein or fragments thereof are useful for improving or reducing symptoms of, or treating or preventing osteoarthritis. See, for example, Malemud CJ. , Cytokines as therapeutic targets for osteoarthritis, BioDrugs,

2004; 18 (1): 23-35; Westacott CI, et al, Cytokines in osteoarthritis: mediators or markers of joint destruction ?,

Semin Arthritis Rheum., 1996 Feb; 25 (4): 254-72; Sugiyama T., Involvement of interleukin-6 and prostaglandin E2 in particular osteoporosis of postmenopausal women with rheumatoid arthritis, J Bone Miner Metab., 2001; 19 (2): 89-96; Abrahamsen B, et al, Cytokines and bone loss in a 5-year longitudinal study hormone replacement therapy suppresses serum soluble interleukin-6 receptor and increases interleukin-l-receptor

447

MEXICAN INSTITUTE OF PROPERTY antagonist: the Danish Osteoporosis Prevent ion ΙΝ§ ^ ΰ! ^ Γ Miner Res., August 2000; 15 (8): 1545-5471 St

Hormone replacement therapy and interrelation between serum interleukin-6 and body mass index in postmenopausal women: a population-based study, J Clin Endocrinol Metab., March 2000; 85 (3): 1340-4; Manolagas SC, The role of IL-6 type cytokines and their receptors in bone, Ann NY Acad Sci., 1998 May 1; 840: 194-204; Ershler WB, et al, Immunologic aspects of osteoporosis, Dev Comp Immunol., 1997 Nov-Dec; 21 (6): 487-99;

Jilka RL, et al, Increased osteoclast development after estrogen loss: mediation by interleukin-6, Science, 1992 Jul 3; 257 (5066): 88-91; Rallen KJ, et al, New developments in IL-6 dependent biology and therapy: where do we stand and what are the options ?, Expert Opin Investig Drugs, September

1999; 8 (9): 1327-49; Neale SD, et al, The influence of serum cytokines and growth factors on osteoclast formation in Paget's disease, QJM, April 2002, -95 (4): 233-40; Roodman GD,

Osteoclast function In Paget's disease and multiple myeloma,

Bone, August 1995, -17 (2 Suppl): 57S-61S; Hoyland JA, et al, Interleukin-6, IL-6 receptor, and IL-6 nuclear factor gene expression in Paget's disease, J Bone Miner Res., January

1994; 9 (1): 75-80; and Roodman GD, et al, Interleukin 6. A potential autocrine / paracrine factor in Paget's disease of bone, J Clin Invest., January 1992; 89 (1): 46-52; the descriptions of which are incorporated herein in their entirety as

448

IMPI Mexican institute.

OF THE PROPERTY

INDUSTRIAL ““ -

reference.

In another embodiment of the invention, the anti-IL-6 antibodies described herein or fragments thereof are useful for improving or reducing symptoms of, or treating or preventing diseases and disorders associated with cancer. Cancer-associated diseases and disorders include but are not limited to acanthoma, acinic cell carcinoma, acoustic neuroma, acral lentiginous melanoma, acrospiroirta, acute eosinophilic leukemia, acute lymphoblastic leukemia, acute megakaryoblastic leukemia, acute monocytic leukemia with maturation, dendritic cell acute myeloid leukemia, acute myeloid leukemia, acute promyelocytic leukemia, adamantinoma, adenocarcinoma, Adenoid cystic carcinoma, adenoma, adenomatoid odontogenic tumor, adrenocortical carcinoma, adult T-cell leukemia, aggressive NK-cell leukemia, AIDS-related cancers, AIDS soft-tissue alveolar sarcoma, ameloblastic fibroma, anal cancer, anaplastic lymphoma large cell, anaplastic thyroid cancer, angioimmunoblastic T-cell lymphoma, angiomyolipoma, angiosarcoma, cancer of the appendix, astrocytoma, atypical rhabdoid teratoid tumor, basal cell carcinoma, basal cell carcinoma, B cell leukemia, B cell lymphoma, Bellini's duct carcinoma, biliary tract cancer, bladder cancer, blastoma, bone cancer, bone tumor glioma

449

MEXICAN INSTITUTE

OF THE PROPERTY VjtaiGrfili INDUSTRIAL

brain stem, brain tumor, breast cancer, tumor

Brenner's, bronchial tumor, bronchioloalveolar carcinoma, Brown's tumor, Burkitt's lymphoma, cancer of unknown primary site, carcinoid tumor, carcinoma, carcinoma in situ, carcinoma of the penis, carcinoma of unknown primary site, carcinosarcoma, Castleman's disease, embryonal tumor of the central nervous system, cerebellar astrocytoma, cerebral astrocytoma, cervical cancer, cholangiocarcinoma, chondroma, chondrosarcoma, chordoma, choriocarcinoma, choroid plexus papilloma, chronic lymphocytic leukemia, chronic monocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disease, chronic neutrophilic leukemia, clear cell tumor, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, disease of

Degos, dermatofibrosarcoma protuberans, dermoid cyst, desmoplastic small round cell tumor, diffuse large B cell lymphoma, dysembryoplastic neuroepithelial tumor, embryonal carcinoma, endodermal sinus tumor, endometrial cancer, endometrial uterine cancer, endometrioid tumor, T cell-associated lymphoma enteropathy, ependymoblastoma, ependymoma, epithelioid sarcoma, erythroleukemia, esophageal cancer, esthesioneuroblastoma, Ewing family tumor, Ewing family sarcoma, Ewing sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer,

450 xvi ri

MEX'CANO INSTITUTE

DE LA HBOHLDAD V, '- ·;

INDUSTRIAL Extramammary Paget's disease, fallopian tube cancer,

Fetus in fetu, fibroma, fibrosarcoma, follicular lymphoma, follicular thyroid cancer, gallbladder cancer, gallbladder cancer, ganglioglioma, ganglioneuroma, gastric cancer, gastric lymphoma, gastrointestinal cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, tumor of the gastrointestinal stroma, germ cell tumor, germinoma, gestational choriocarcinoma, gestational trophoblastic tumor, giant cell bone tumor, glioblastoma multiforme, glioma, gliomatosis cerebri, glomus tumor, glucagonoma, gonadoblastoma, granule cell tumor, hairy cell leukemia, hairy cell leukemia, head and neck cancer, head and neck cancer, heart cancer, hemangioblastoma, hemangiopericytoma, hemangiosarcoma, malignancy hematologic, hepatocellular carcinoma, hepatosplenic T-cell lymphoma, hereditary ovarian and breast cancer syndrome, Hodgkin lymphoma, Hodgkin lymphoma, hypopharyngeal cancer, hypothalamic glioma, inflammatory breast cancer, intraocular melanoma, islet cell carcinoma, islet cell tumor, juvenile myelomonocytic leukemia, Kaposi sarcoma, Kaposi's sarcoma, kidney cancer, Klatskin tumor, Krukenberg tumor, cancer of larynx, laryngeal cancer, lentigo maligna melanoma, leukemia, leukemia, cancer of the lip and oral cavity, liposarcoma, lung cancer, luteoma,

451

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MSXiCANO DELA PÍ.OHICAO INSTITUTE

INDUSTRIAL lymphangioma, lymphangiosarcoma, lymphoepithelioma, lymphoid leukemia, lymphoma, macroglobulinemia, fibrous malignant histiocytoma, malignant fibrous histiocytoma, malignant fibrous histiocytoma, malignant neoplasm, malignant neural tumor, malignant tumor, , mantle cell lymphoma, mast cell leukemia, mediastinal germ cell tumor, mediastinal tumor, medullary thyroid cancer, medulloblastoma, medulloblastoma, medulloepithelioma, melanoma, melanoma, meningioma, Merkel cell carcinoma, mesothelioma, mesothelioma, metastatic squamous neck cancer with occult primary tumor, mixed urothelial carcinoma, mixed Müllerian tumor, monocytic leukemia, mouth cancer, mucinous tumor, syndrome multiple endocrine neoplasia, multiple myeloma, multiple myeloma, mycosis fungoides, mycosis fungoides, myelodysplastic disease, myelodysplastic syndromes, myeloid leukemia, myeloid sarcoma, myeloproliferative disease, myxoma, cancer of the nasal cavity, nasopharyngeal cancer, nasopharyngeal carcinoma, neoplasm, neurinoma, neuroblastoma, neuroblastoma, neurofibroma, neuroma, nodular melanoma, non-Hodgkin lymphoma, non-melanoma skin cancer, non-small cell lung, ocular oncology, oligoastrocytoma, oligodendroglioma, oncocytoma, meningioma of the optic nerve sheath, oral cancer, oral cancer, oropharyngeal cancer,

452

IMPI

MEXICAN PROPERTY INSTITUTE

INDUSTRIAL osteosarcoma, osteosarcoma, ovarian cancer, ovarian cancer, epithelial ovarian cancer, ovarian germ cell tumor, low malignant potential ovarian tumor, Paget disease of the breast, Pancoast tumor, pancreatic cancer, pancreatic cancer, papillary thyroid cancer, papillomatosis, paraganglioma, paranasal sinus cancer, parathyroid cancer, penile cancer, perivascular epithelioid cell tumor, pharyngeal cancer, pheochromocytoma, Pineal parenchymal tumor of intermediate differentiation, pineoblastoma, pituicytoma, pituitary adenoma, pituitary tumor, plasma cell neoplasm, pleuropulmonary blastoma, polymeryoma, lymphoblastic precursor lymphoma of T cells, primary lymphoma of the central nervous system, primary effusion lymphoma, primary liver cancer, primary peritoneal cancer, primitive neuroectodermal tumor, prostate cancer, peritoneal pseudomyxoma, rectal cancer, Renal cell carcinoma, NUT gene related respiratory tract carcinoma on chromosome 15, retinoblastoma, rhabdomyoma, rhabdomyosarcoma, Richter transformation, sacrococcygeal teratoma, cancer of the salivary gland, sarcoma, Schwannomatosis, sebaceous gland carcinoma, neoplasm seminoma, serous tumor, Sertoli-Leydig cell tumor, sex cord stromal tumor, Sézary syndrome, signet ring cell carcinoma, skin cancer, small round blue cell tumor

453

ΙΜ

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY small cell carcinoma, microcytic lung cancer, small cell lymphoma, cancer of the small intestine, soft tissue sarcoma, somatostatinoma, chimney sweep wart, spinal cord tumor, spinal tumor, zone lymphoma marginal splenic, squamous cell carcinoma, stomach cancer, disseminating superficial melanoma, primitive supratentorial neuroectodermal tumor, epitalial ovarian tumor, synovial sarcoma, acute T-cell leukemia, large granular T-cell lymphocytic leukemia, T-cell leukemia, T-cell lymphoma, T-cell prolymphocytic leukemia, teratoma, terminal lymphatic cancer, testicular cancer, thecoma, throat cancer, thymic carcinoma, thymoma, thyroid cancer, transitional cell cancer of the renal pelvis and ureter, transitional cell carcinoma, urachal cancer, urethral cancer, urogenital neoplasm, uterine sarcoma, uveal melanoma, vaginal cancer, Verner Morrison syndrome, warty carcinoma, visual glioma, vulvar cancer, Waldenstrom's macroglobulinemia, Warthin's tumor, Wilms' tumor, or any combination thereof as well as drug resistance in cancer chemotherapy and chemotherapy toxicity against cancer. See, for example, Hirata T, et al, Humanized antiinterleukin-6 monoclonal receptor antibody induced apoptosis of fresh and cloned human myeloma cells in vitro, Leuk Res., April 2003; 27 (4): 343-9, Bataille R, et al, Biologic effects of

454

advanced

INSTITUTO MCX'CANO DE LA FROPitL A3

INDUSTRIAD anti-interleukin-6 murine monoclonal antibody in multiple myeloma, Blood, July 15, 1995; 86 (2): 685-91;

Goto H, et al, Mouse anti-human interleukin-6 monoclonal receptor antibody inhibits proliferation of fresh human myeloma cells in vitro, Jpn J Cáncer Res., September

1994; 85 (9): 958-65; Klein B, et al, Murine anti-interleukin-6 monoclonal antibody therapy for a patient with plasma cell leukemia, Blood, September 1, 1991; 78 (5): 1198-204; Mauray S, et al, I Epstein-Barr virus-dependent lymphoproliferative disease: critical role of IL-6, Eur J Immunol., July 2000; 30 (7): 2065-73; Tsunenari T, et al, New xenograft model of multiple myeloma and efficacy of a humanized antibody against human interleukin-6 receptor, Blood, September 15, 1997; 90 (6): 2437-44; Emilie D, et al, Interleukin-6 production in high-grade B lymphomas: correlation with the presence of malignant immunoblasts in acquired immunodeficiency syndrome and in human immunodeficiency virus-seronegative patients, Blood, July 15, 1992; 80 (2): 498-504; Emilie D, et al, Administration of an anti-interleukin-6 monoclonal antibody to patients with acquired immunodeficiency syndrome and lymphoma: effect on lymphoma growth and on B clinical Symptoms, Blood, October 15, 1994; 84 (8): 2472-9; Smith PC, et al, Antiinterleukin-6 monoclonal antibody induces regression of human prostate cancer xenografts in nude mice, Prostate, June 15, 2001; 48 (1): 47-53; Smith PC, et al, Interleukin-6 and

455

IMPI

McXICANO DE LA PROPERTY INDUSTRIAL prostate cancer progression, Cytokine Growth Factor Rev., March 2001; 12 (1): 33-40; Chung TD, et al, Characterization of the role of IL-6 in the progression of prostate cancer, Prostate, February 1999; 38 (3): 199-207; Okamoto M, et al,

Interleukin-6 as a paracrine and autocrine growth factor in human prostatic carcinoma cells in vitro, Cancer Res., 1997 Jan

1; 57 (1): 141-6; Reittie JE, et al, Interleukin-6 inhibits apoptosis and tumor necrosis factor induced proliferation of Bchronic lymphocytic leukemia, Leuk Lymphoma, June 1996; 22 (l-2): 83-90, follow 186, piate color VI; Sugiyama H, et al, The expression of IL-6 and its related genes in acute leukemia, Leuk Lymphoma, March 1996; 21 (1-2): 49-52; Bataille

R, et al, Effects of an anti-interleukin-6 (IL-6) murine monoclonal antibody in a patient with acute monoblastic leukemia, Med Oncol Tumor Pharmacother., 1993; 10 (4): 185-8;

Kedar I, et al, Thalidomide reduces serum C-reactive protein and interleukin-6 and induces response to IL-2 in a fraction of metastatic renal cell cancer patients who failed IL-2-based therapy, Int J Cáncer, 2004 Jun 10; 110 (2): 260-5; Angelo LS, Talpaz M, Kurzrock R, Autocrine interleukin-6 production in renal cell carcinoma: evidence for the involvement of p53, Cáncer Res., 2002 Feb 1; 62 (3): 932-40; Nishimoto N, Humanized anti-interleukin-6 receptor antibody treatment of multicentric Castleman disease, Blood, 2005 Oct 15; 106 (8): 2627-32, Epub 2005 Jul 5; Katsume A, et al, Anti-interleukin 6 (IL-6) receptor

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MEXICAN INSTITUTE CS THE PROPERTY

INDUSTRIAL

antibody suppresses Castleman's disease like symptoms emerged in IL-6 transgenic mice, Cytokine, 2002 Dec 21; 20 (6): 304-11; Nishimoto N, et al, Improvement in Castleman's disease by humanized anti-interleukin-6 receptor antibody therapy, Blood,

2000 Jan 1; 95 (1): 56-61; Screpanti I, Inactivation of the IL-6 gene prevents development of multicentric Castleman's disease in C / EBP beta-deficient mice, J Exp Med., 1996 Oct 1; 184 (4): 1561-6; Hsu SM, et al, Expression of interleukin-6 in

Castleman's disease, Hum Pathol., 1993 Aug; 24 (8): 833-9;

Yoshizaki K, et al, Pathogenic significance of interleukin-6 (IL 6 / BSF-2) in Castleman's disease, Blood, 1989 Sep; 74 (4): 1360-7; Nilsson MB, et al, Interleukin-6, secreted by human ovarian carcinoma cells, is a potent proangiogenic cytokine, Cancer Res., 2005 Dec 1; 65 (23): 10794-800; Toutirais

O, et al, Constitutive expression of TGF-betal, interleukin-6 and interleukin-8 by tumor cells as a major component of immune escape in human ovarian carcinoma, Eur Cytokine Netw., 2003

Oct-Dec; 14 (4): 246-55; Obata NH, et al, Effects of interleukin 6 on in vitro cell attachment, migration and invasion of human ovarian carcinoma, Anticancer Res., 1997 Jan-Feb; 17 (1A): 33742; Dedoussis GV, et al, Endogenous interleukin 6 conveys resistance to cis-diamminedichloroplatinum-mediated apoptosis of the K562 human leukemic cell line, Exp Cell Res., 1999 Jun 15; 249 (2): 269-78; Borsellino N, et al, Blocking signaling through the Gpl30 receptor chain by interleukin-6 and

457

INSTITUI O MEXICANO

OF PROPERTY V,

INDUSTRIAL oncostatin M inhibits PC-3 cell growth and sensitizes the tumor cells to etoposide and cisplatin-mediated cytotoxicity, Cancer, 1999 Jan 1; 85 (1): 134-44; Borsellino N, et al, Endogenous interleukin 6 is a resistance factor for cisdiamminedichloroplatinum and etoposide-mediated cytotoxicity of human prostate carcinoma cell lines, Cáncer Res., 1995 Oct

15; 55 (20): 4633-9; Mizutani Y, et al, Sensitization of human renal cell carcinoma cells to cis-diamminedichioroplatinum (II) by anti-interleukin 6 monoclonal antibody or anti-interleukin 6 receptor monoclonal. antibody; Cancer Res., 1995 Feb

1; 55 (3): 590-6; Yusuf RZ, et al, Paclitaxel resistance:

molecular mechanisms and pharmacologic manipulation, Curr Cancer Drug Targets, 2003 Feb; 3 (1): 1-19; Duan Z, et al,

Overexpression of IL-6 but not IL-8 increases paclitaxel resistance of U-20S human osteosarcoma cells, Cytokine, 2002

Mar 7; 17 (5): 234-42; Conze D, et al, Autocrine production of interleukin 6 causes multidrug resistance in breast cancer cel ls, Cancer Res., 2001 Dec 15; 61 (24): 8851-8; Rossi JF, et al,

Optimizing the use of anti-interleukin-6 monoclonal antibody with dexamethasone and 140 mg / m2 of melphalan in multiple myeloma: results of a pilot study including biological aspects, Bone Marrow Transplant, 2005 Nov; 36 (9): 771-9; and Tonini G, et al, Oxaliplatin may induce cytokine-release syndrome in colorectal cancer patients, J Biol Regul Homeost Agents, 2002 Apr-Jun; 16 (2): 105-9; whose descriptions are incorporated in the

458

IMPI

MEXICAN INSTITUTE OF LA PRORI5DAL> INDUSTRIAL present in its entirety as a reference.

In another embodiment of the invention, the anti-IL-6 antibodies described herein or fragments thereof are useful for improving or reducing symptoms of, or treating or preventing ischemic heart disease, atherosclerosis, obesity, diabetes, asthma, multiple sclerosis, Alzheimer's disease, cerebrovascular disease, fever, acute phase response, allergies, anemia, anemia of inflammation (anemia of chronic disease), hypertension, depression, depression associated with chronic disease, thrombosis, thrombocytosis, acute heart failure, metabolic syndrome, miscarriages, obesity, chronic prostatitis, glomerulonephritis, pelvic inflammatory disease, reperfusion injury, and transplant rejection. See, for example, Tzoulaki I, et al,

C-reactive protein, interleukin-6, and soluble adhesion molecules as predictors of Progressive peripheral atherosclerosis in the general population: Edinburgh Artery Study, Circulation, 2005 Aug 16; 112 (7): 976-83, Epub 2005 Aug 8; Rattazzi M, et al, C-reactive protein and interleukin-6 in vascular disease: culprits or passive bystanders ?, J

Hypertens., 2003 Oct; 21 (10): 1787-803; Ito T, et al, HMG-CoA reductase inhibitors reduce interleukin-6 synthesis in human vascular smooth muscle cells, Cardiovasc Drugs Ther., 2002 Mar; 16 (2): 121-6; Stenvinkel P, et al, Mortality, malnutrition( and atherosclerosis in ESRD: what is the role of interleukin459

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

6 ?, Kidney Int Suppl., 2002 May; (80): 103-8; Yudkin JS, et al, Inflammation, obesity, stress and coronary heart disease: is interleukin-6 the link ?, Atherosclerosis, February

2000; 148 (2). * 209-14; Huber SA, et al, Interleukin-6 exacerbates early atherosclerosis in mice, Arterioscler Thromb Vaso Biol., October 1999; 19 (10): 2364-7; Kado S, et al, Circulating levels of interleukin-6, its soluble receptor and interleukin6 / interleukin-6 receptor complexes in patients with type 2 diabetes mellitus, Acta Diabetol., June 1999; 36 (1-2): 67-72 ;

Sukovich DA, et al, Expression of interleukin-6 in atherosclerotic lesions of male ApoE-knockout mice: inhibition by 17beta-estradiol, Arterioscler Thromb Vasc Biol., 1998 Sept; 8 (9): 1498-505, * Klover PJ, et al, Interleukin-6 depletion selectively improves hepatic insulin action in obesity,

Endocrinology, 2005 Aug; 146 (8). * 3417-27, Epub 2005 Apr 21; read

YH, et al, The evolving role of inflammation in obesity and the metabolic syndrome, Curr Diab Rep., 2005 Feb; 5 (1). * 70-5; Diamant

M, et al, The association between abdominal visceral fat and carotid stiffness is mediated by circulating inflammatory 20 markers in uncomplicated type 2 diabetes, J Clin Endocrinol

Metab., 2005 Mar; 90 (3): 1495-501, Epub 2004 Dec 21; Bray GA, Medical consequences of obesity, J Clin Endocrinol Metab., 2004 Jun; 89 (6): 2583 9; Klover PJ, et al, Chronic exposure to interleukin-6 causes hepatic insulin resistance in mice,

Diabetes, 2003 Nov; 52 (ll): 2784-9; Yudkin JS, et al,

460

Inflammation, obesity, stress and coronary heart disease: is interleukin-6 the link ?, Atherosclerosis, 2000 Feb; 148 (2); 20914; Doganci A, et al, Pathological role of IL-6 in the experimental allergic bronchial asthma in mice, Clin Rev

Allergy Immunol., 2005 Jun; 28 (3): 257-70; Doganci A, et al, The

IL-6R alpha chain Controls lung CD4 + CD25 + Treg development and function during allergic airway inflammation in vivo, J Clin

Invest., 2005 Feb; 115 (2); 313 25, (Erratum in: J Clin Invest.,

2005 May; '115 (5) :: 1388, Lehr, Hans A [added]); Stelmasiak Z, et al, IL 6 and SIL-6R concentration in the cerebrospinal fluid and serum of MS patients, Med Sci Monit., 2001 SepOct; 7 (5): 914-8; Tilgner J, et al, Continuous interleukin-6 application in vivo via macroencapsulation of interleukin-6expressing COS-7 cells induces massive gliosis, Glia, 2001

Sep; 35 (3): 234-45, Brunello AGr et al, Astrocytic alterations in interleukin-6 Soluble interleukin-6 receptor alpha doubletransgenic mice, Am J Pathol., 2000 Nov; 157 (5): 1485-93; Hampel H, et al, Pattern of interleukin-6 receptor complex immunoreactivity between cortical regions of rapid autopsy normal and Alzheimer's disease brain, Eur Arch Psychiatry Clin Neurosci., 2005 Aug; 255 (4): 269-78, Epub 2004 Nov 26; Cacquevel M, et al, Cytokines in neuroinflammation and Alzheimer's disease, Curr Drug Targets, 2004 Aug; 5 (6): 529-34; Quintanilla RA, et al, Interleukin 6 induces Alzheimer-type phosphorylation of tau protein by deregulating the cdk5 / p35 pathway, Exp Cell

461

Res., 2004 Apr 15; 295 (1): 245-57; Gadient

MEXICAN INSTITUTE OF LA?. <O? IFDAL>

industrial

Interleukin-6 (IL-6) —a molecule with both beneficial and destructive potentials, Prog Neurobiol., 1997 Aug; 52 (5): 379-90;

Hull M, et al, Occurrence of interleukin-6 in cortical plaques of Alzheimer's disease patients may precede transformation of diffuse into neuritic plaques, Ann NY Acad Sci., 1996 Jan

17; 777: 205-12; Rallidis LS, et al, Inflammatory markers and inhospital mortality in acute ischemic stroke, Atherosclerosis,

2005 Gave 30; Emsley HC, et al, Interleukin-6 and acute ischemic stroke, Acta Neurol Scand., 2005 Oct; 112 (4): 273-4;

Smith CJ, et al, Peak plasma interleukin-6 and other peripheral markers of inflammation in the first week of ischaemic stroke correlate with brain infarct volume, stroke severity and longterm outcome, BMC Neurol., 2004 Jan 15; 4: 2; Vila N, et al,

Proinflammatory cytokines and early neurological worsening in ischemic stroke, Stroke, 2000 Oct; 31 (10): 2325-9; and Tarkowski

E, et al, Early intrathecal production of interleukin-6 predicts the size of brain injury in stroke, Stroke, 1995

Aug; 26 (8): 1393-8; the descriptions of which are incorporated herein by reference in their entirety.

462

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MEXICAN INSTITUTE -, ---. 2 Λί

KOI-IEDAD

OF THE PKOI'I

INDUSTRIAL cytokine crises include, but are not limited to, graft versus host disease (GVHD), avian influenza, smallpox, pandemic influenza, adult respiratory distress syndrome (ARDS), severe acute respiratory syndrome (SARS), sepsis, and systemic inflammatory response (SIRS). See for example

Cecil, RL, Goldman, L., & Bennett, JC (2000). Cecil textbook of medicine. Philadelphia: WB Saunders; Ferrara JL, et al., Cytokine storm of graft-versus-host disease: a critical effector role for interleukin-1, Transplant Proc. 1993 Feb; 25 (l

Pt 2): 1216-7; Osterholm MT, Preparing for the Next Pandemic, N Engl J Med. 2005 May 5; 352 (18): 1839-42; Huang KJ, et al., An interferon-gamma-related cytokine storm in SARS patients, J Med Virol. 2005 Feb; 75 (2): 185-94; and Cheung CY, et al., Induction of proinflammatory cytokines in human macrophages by influenza A (H5N1) viruses: a mechanism for the unusual severity of human disease? Lancet. 2002 Dec 7; 360 (9348): 1831-7.

In another embodiment of the invention, the anti-IL-6 antibodies described herein or fragments thereof, are useful as an insomnia aid.

Administration

In one embodiment of the invention, the anti-IL-6 antibodies described herein or IL-6 binding fragments thereof, as well as combinations of such antibody fragments, are administered to a subject in a concentration of

463

between about 0.1 and 20 mg / kg, such as about 0.4 mg / kg, about 0.8 mg / kg, about T? 6 mg / kg, or about 4 mg / kg, of the body weight of the recipient subject. In a preferred embodiment of the invention, the anti-IL-6 antibodies described herein or IL-6 binding fragments thereof, as well as combinations of such antibody fragments, are administered to a subject in a concentration of about 0.4 mg / kg of the body weight of the recipient subject. In a preferred embodiment of the invention, the anti-IL-6 antibodies described herein or IL-6 binding fragments thereof, as well as combinations of such antibody fragments, are administered to a recipient subject with a frequency of once every twenty-six weeks or less, such as once every sixteen weeks or less, once every eight weeks or less, or once every four weeks or less. In another preferred embodiment of the invention, the anti-IL-6 antibodies described herein or IL-6 binding fragments thereof, as well as combinations thereof, are administered to a recipient subject with a frequency of at most once during a period of approximately one week, such as, at most once during a period of approximately two weeks such as, at most once during a period of approximately four weeks, such as, at most once during a period of approximately eight weeks, such as, at most once during a period of approximately twelve

464

IMPIAS

MEXICAN INSTITUTE

OF PROPERTY C-dj.

INDUSTRIAL '«YOUR weeks, such as, at most once during a period of approximately sixteen weeks, such as, at most once during a period of approximately twenty-four weeks.

It is understood that the effective dose will depend on the attributes of the recipient subject, such as, for example, age, sex, state of pregnancy, body mass index, lean body mass, condition or conditions for which the composition is provided, other conditions conditions of the recipient subject that may affect the metabolism or tolerance of the composition, levels of IL-6 in the recipient subject and resistance to the composition (for example, that arise when the patient develops antibodies against the composition). A person skilled in the art will be able to determine an effective dose and frequency of administration through routine experiments, for example, guided by the descriptions herein and the teachings in Goodman, LS, Gilman, A.,

Brunton, LL, Lazo, JS, & Parker, KL (2006). Goodman &

Gilman's the pharmacological basis of therapeutics. NY:

McGraw-Hill; Howland, RD, Mycek, MJ, Harvey, RA,

Champe, PC, & Mycek, MJ (2006). Pharmacology.

Lippincott's illustrated reviews. Philadelphia: Lippincott Williams & Wilkins; and Golan, DE (2008). Principles of pharmacology: the pathophysiologic basis of drug therapy.

Philadelphia, Pa., [Etc.]: Lippincott Williams & Wilkins.

In another embodiment of the invention, anti465 antibodies

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MEXICAN INSTITUTE DS LA PROVICCAD INDUSIRiAL

IL-6 described herein or the IL-6 binding fragments thereof, as well as combinations of such antibody fragments, are administered to a subject as a pharmaceutical formulation.

A pharmaceutical composition refers to a chemical or biological composition suitable for administration to a mammal. Such compositions can be specifically formulated for administration by one or more routes, including but not limited to buccal, epicutaneous, epidural, inhalation, intraarterial, intracardiac, intracerebroventricular, intradermal, intramuscular, intranasal, infraocular, intraperitoneal, intraspinal, intrathecal, intravenous , oral, parenteral, rectal by means of an enema or suppository, subcutaneous, subdermal, sublingual, transdermal and transmucosal. Furthermore, administration can occur by injection means, powder, liquid, gel, drops, or other means of administration.

In an embodiment of the invention, the anti-IL-6 antibodies described herein, or the IL-6 binding fragments thereof, as well as combinations of such antibody fragments, can optionally be administered in combination with one or more agents assets. Such active agents include analgesic, antipyretic, anti-inflammatory, antibiotic, antiviral and anticytokine agents. Active agents include TNF-α, IL-2 agonists, antagonists, and modulators,

466

IMPI

INDUSTRIAL

IL-4, IL-6, IL-10, IL-12, IL-13, IL-18, IFN-a, IFN-γ, BAFF,

CXCL13, IP-10, VEGF, EPO, EGF, HRG, hepatocyte growth factor (HGF), Hepcidin, including antibodies reactive against any of the above and antibodies reactive against any of its receptors. Active agents also include 2-Arylpropionic acids, Aceclofenac,

Acemetacin, Acetylsalicylic Acid (Aspirin), Alclofenac, Alminoprofen, Amoxiprine, Ampirone, Arilalkanoic Acids,

Azapropazone, Benorylate / Benorylate, Benoxaprofen, Bromfenac, 10 Carprofen, Celecoxib, Choline and Magnesium Salicylate,

Clofezone, COX-2 inhibitors, Dexibuprofen, Dexketoprofen, Diclofenac, Diflunisal, Droxicam, Etenzamide, Etodolac,

Etoricoxib, Faislamine, Phenic Acids, Fenbufen, Fenoprofen, Flufenamic Acid, Flunoxaprofen, Flurbiprofen,

Ibuprofen, Ibuproxam, Indomethacin, Indoprofen, Kebuzone,

Ketoprofen, Ketorolac, Lornoxicam, Loxoprofen, Lumiracoxib, Magnesium Salicylate, Meclofenamic Acid, Mefenamic Acid,

Meloxicam, Metamizole, Methyl salicylate, Mofebutazone,

Nabumetone, Naproxen, N-arylanthranilic acids, Oxametacin, 20 Oxaprozin, Oxicams, Oxyphenbutazone, Parecoxib, Phenazone,

Phenylbutazone, Phenylbutazone, Piroxicam, Pirprofen, Profenos, Proglumetacina, derivatives of Pirazolidina, Rofecoxib, salicylate of

Sulfinpyrazone,

Salicyl,

Sulindac,

Salicylamide, Salicylates, Suprofen, Tenoxicam, acid

Tiaprófenico, Tolfenamic Acid, Tolmetin and Valdecoxib. The

467

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MEXICAN INSTITUTE

D £ PROPERTY V? = Huh £ ¡LV »3. ,. ,, - „. ,, „. ,. . INDUSTRIAL r.

Antibiotics include Amikacin, Ammoglycosides, Amoxicillma,

Dalfopristina,

Ampicillin, Ansamycins, Arsphenamine, Azithromycin, Azlocilin, Aztreonam, Bacitracin, Carbacefem, Carbapenems, Carbenicillin,

Cefaclor, Cefadroxil, Cefalexina, Cefalotina, Cefalotina,

Cefamandol, Cefazolin, Cefdinir, Cefditoren, Cefepime, Cefixime, Cefoperazone, Cefotaxime, Cefoxitin, Cefpodoxime,

Cefprozil, Ceftazidime, Ceftibuten, Ceftizoxime, Ceftobiprol,

Ceftriaxone, Cefuroxime,. Cephalosporinsa, Chloramphenicol ·,

Cilastatin, Ciprofloxacin, Clarithromycin, Clindamycin,

Chloxacillin, Colistin, Co-trimoxazole,

Demeclocycline, Dicloxacillin, Dirithromycin, Doripenem, Doxycycline, Enoxacin, Ertapenem, Erythromycin, Ethambutol, Flucloxacillin, Phosphomycin, Furazolidone, Fusidic acid, Gatifloxacin, Geldanamicin, Gentamycin, Glycopeptides,

Herbimycin, Imipenem, Isoniazid, Kanamycin, Levofloxacin,

Lincomycin, Linezolid, Lomefloxacin, Loracarbef, Macrolides,

Mafenide, Meropenem, Methicillin, Metronidazole, Mezlocillin,

Minocycline, Monobactams, Moxifloxacin, Mupirocin, Nafcillin,

Neomycin, Netilmicin, Nitrofurantoin, Norfloxacin,

Oxacillin, Oxytetracycline, Paromomycin,

Ofloxacin,

Penicillin,

Polymyxin

Quinolones,

Penicillins, Piperacillin, Platensimycin, B, Polypeptides, Prontosil, Pyrazinamide, Quinupristin, Rifampin, Rifampin,

Roxithromycin, Spectinomycin, Streptomycin, Sulfacetamide,

Sulfametizole, Sulfanilimide, Sulfasalazine, Sulfisoxazole,

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Sulfonamides,

Tetracyclines,

Trimethoprim,

MEXICAN INSTITUTE OF PROPERTY

Teicoplanin, Telithromycin, , NDJS'[fe't

Tinidazoi,

TóbrarnTciña, '

Ticarcillin,

Trimethoprim-Sulfamethoxazole, Troleandomycin, Trovafloxacin and Vancomycin. Active agents also include Aldosterone, Beclomethasone, Betamethasone,

Corticosteroids, Cortisol, Cortisone acetate, acetate

Deoxycorticosterone, Dexamethasone, Fludrocortisone Acetate, Glucocorticoids, Hydrocortisone, Methylprednisolone, Prednisolone, Prednisone, Spheroids, and Triamcinolone. Antiviral agents include abacavir, acyclovir, acyclovir, adefovir, amantadine, amprenavir, a fixed-dose combination antiretroviral, a synergistic antiretroviral enhancer, arbidol, atazanavir, atripla, brivudine, cidofovir, combivir, darunavir, delavirdine, didanos efavirenz, emtricitabine, enfuvirtide, entecavir, entry inhibitors, famciclovir, fomivirsen, fosamprenavir, foscarnet, fosonet, fusion inhibitor, ganciclovir, gardasil, ibacitabine, idoxuridine, imiquimod, imunovir, indinavir, inosine, integrase inhibitor, interferon, interferon type I, interferon type II, interferon type III, lamivudine, lopinavir, loviride, wonderiroc, MK-0518, moroxidine, nelfinavir, nevirapine, nexavir, analogues of nucleoside, oseltamivir, penciclovir, peramivir, pleconaryl, podophyllotoxin, protease inhibitor, reverse transcriptase inhibitor, ribavirin, rimantadine, ritonavir, saquinavir, stavudine, tenofovir,

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trizivir,

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY tenofovir disoproxil, tipranavir, trifluridine, tromantadine, truvada, valaciclovir, valganciclovir, vicriviroc, vidarabine, viramidine, zalcitabine, zanamivir and zidovudine. Any suitable combination of these active agents is also contemplated.

A pharmaceutical excipient or a pharmaceutically acceptable excipient is a carrier, usually a liquid, where an active therapeutic agent is formulated. In an embodiment of the invention, the therapeutically active agent is a humanized antibody described herein, or one or more fragments thereof. The excipient generally does not provide any pharmacological activity to the formulation although it can provide chemical and / or biological stability and release characteristics. Examples of formulations can be found, for example, in Remington's Pharmaceutical Sciences, 19 * Ed., Grennaro, A., Ed., 1995, which is incorporated herein by reference.

As used herein a pharmaceutically acceptable carrier or excipient includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption retarding agents that are physiologically compatible. In one embodiment, the carrier is suitable for parenteral administration. Alternatively, the carrier may be suitable for intravenous administration,

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intraperitoneal, intramuscular or sublingual. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional medium or agent is incompatible with the active compound, the use thereof is contemplated in the pharmaceutical compositions of the invention.

Complementary active compounds can also be incorporated into the compositions.

In an embodiment of the invention, these can be used to administer antibodies of the invention intravenously, including Abl, for indications in cancer; the administration formulation comprises, or alternatively consists of, about 10.5 mg / mL antibody, 25 mM histidine base, pH 6 qsp phosphoric acid and 250 mM sorbitol.

In another embodiment of the invention, these can be used to administer antibodies of the invention intravenously, including Abl, for indications in cancer; the administration formulation comprises, or alternatively consists of, about 10.5 mg / mL antibody, 12.5 mM histidine base, 12.5 mM histidine hydrochloride (or 25 mM histidine base and hydrochloric acid

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qs at pH 6),

250 Sorbitol mM

IN DU J'í AL v

0.015% (w / w) Polysorbate 80.

In an embodiment of the invention that can be used to administer antibodies of the invention subcutaneously, including Abl, for indications in rheumatoid arthritis, the administration formulation comprises, or alternatively consists of, about 50 or 100 mg / mL of antibody, about 5mM histidine base, about 5mM HC1 histidine to achieve a final pH of 6.250mM sorbitol and 0.015% (w / w) polysorbate 80.

In another embodiment of the invention that can be used to administer antibodies of the invention subcutaneously, including Abl, for indications in rheumatoid arthritis, the administration formulation comprises, or alternatively consists of, about 20 or 100 mg / mL of antibody, about 5mM histidine base, about 5mM histidine HC1 to achieve a final pH of 6, 250 at 280 mM sorbitol (or sorbitol in combination with sucrose) and 0.015% (w / w) polysorbate 80, this formulation has a nitrogen free space in the shipping containers.

Pharmaceutical compositions typically must be sterile and stable under manufacturing and storage conditions. The invention contemplates that the pharmaceutical composition is present in a lyophilized form. The composition can be formulated as a solution, a microemulsion,

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IMPI ^

MEXICAN INSTITUTE

OF PROPERTY V> s = K · —Các

INDUSTRIAL ^ "S" liposome or other ordered structure suitable for high drug concentrations. The carrier may be a Solvent or dispersion medium containing, for example, water, ethanol, polyol (eg, glycerol, propylene glycol, and liquid polyethylene glycol) and suitable mixtures thereof. The invention further contemplates the inclusion of a stabilizer in the pharmaceutical composition.

In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of injectable compositions can be caused by the inclusion in the composition of an agent that delays absorption, for example, monostearate and gelatin salts. Furthermore, the alkaline polypeptide can be formulated in a time release formulation, for example in a composition that includes a slow release polymer. Active compounds can be prepared with carriers that will protect the compound from rapid release, such as a controlled release formulation that includes implants and microencapsulated release systems. Biodegradable and biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid, and polylactic and polyglycolic copolymers (PLG). Many methods for the preparation of such formulations are known to those

473

MEXICAN INSTITUTE

DE LA ΓΚΟΛΕϋΑΟ V · - MS? rtK IKOUSnUAL experts in the art.

For each of the described modalities, the compounds can be administered by a variety of dosage forms. Any biologically acceptable dosage form known to those skilled in the art and combinations thereof are contemplated. Examples of such dosage forms include, but are not limited to, reconstitutable powders, elixirs, liquids, solutions, suspensions, emulsions, powders, granules, particles, microparticles, dispersible granules, seals, inhalants, aerosol inhalants, patches, inhalants in particles, implants, reservoir implants, injectables (including subcutaneous, intramuscular, intravenous, and intradermal), infusions, and combinations thereof.

The foregoing description of various illustrated embodiments of the invention is not intended to be exhaustive or to limit the invention to the specific form described. While specific embodiments and examples of the invention are described herein for illustrative purposes, it is possible to make several equivalent modifications within the scope of the invention, as will be recognized by those skilled in the corresponding art. The teachings of the invention provided herein may be applied for purposes other than the examples described above.

These and other changes can be made to the invention.

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INSTITUTO MEXICa.no DE LA PROPIEDAD in view of the detailed description mentioned above-^ In general, in the claims that follow, the terms used should not be understood as limiting the invention to the specific modalities described in the specification and in the claims. Accordingly, its description does not limit the invention but instead the scope of the invention will be fully determined by the claims below.

The invention may be practiced in ways other than those described particularly in the foregoing description and examples. Numerous modifications and variations of the invention are possible in view of the above teachings and are therefore within the scope of the appended claims.

Certain teachings related to methods for obtaining an antigen-specific clonal population of B lymphocytes were described in US Provisional Patent Application No. 60 / 801,412 filed May 19, 2006, the disclosure of which is incorporated herein in its entirety as reference.

Certain teachings related to the humanization of rabbit-derived monoclonal antibodies and preferred sequence modifications to maintain antigen-binding affinity were described in International Application No. 12 / 124,723, corresponding to File No.

475

Antibodv τ

«Λ

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY representative 67858.704001, titled Novel Rabbit

Humanization Method and Humanized Rabbit Antibodies, filed May 21, 2008, the disclosure of which is incorporated herein in its entirety by reference.

Certain teachings related to the production of antibodies or fragments thereof using mating capable yeast and corresponding methods were described in US Patent Application No. 11 / 429,053 filed on May 8, 2006, (US Patent Application Publication No. No. US2006 / 0270045), the disclosure of which is incorporated herein by reference in its entirety.

Certain teachings related to anti-IL6 antibodies, methods for the production of antibodies or fragments thereof using mating capable yeast, and corresponding methods were described in US Provisional Patent Application No. 60 / 924,550 filed on May 21, 2007, the Description is incorporated herein in its entirety by reference.

Certain teachings related to anti-IL-6 antibodies and methods for using those antibodies or fragments thereof to treat cachexia, weakness, fatigue and / or fever were described in filed US Provisional Patent Application No. 61 / 117,839 filed. on November 25,

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ΜΡΙ

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

2008, the description of which is incorporated herein in its entirety by reference.

Certain anti-IL-6 antibody poiinucleotides and polypeptides are described in the sequence listing that accompanies this patent application filing and their disclosure is incorporated herein by reference in its entirety.

The entire description of each cited document (including patents, patent applications, newspaper articles, abstracts, manuals, books, or other descriptions) in Background of the Invention, Detailed Description and Examples, is incorporated herein in its entirety as reference.

The following examples are intended to provide those skilled in the art with a complete description of how to make and use the subject invention and are not intended to limit the scope of what is understood by the invention. Efforts have been made to ensure accuracy with respect to the figures used (eg, quantities, temperatures, concentrations, etc.) but certain errors and experimental deviations should be taken into account. Unless otherwise noted, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees centigrade, and pressure is at or near atmospheric.

EXAMPLES

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Example 1 Production of cultures enriched with antigen-specific B lymphocyte antibodies

Antibody panels are obtained by immunizing traditional antibody host animals to exploit the native immune response to a target antigen of interest. Typically, the host used for immunization is a rabbit or other host that produces antibodies using a similar maturation process and provides a population of antigen-specific B lymphocytes producing antibodies of comparable diversity, eg. epitopic diversity. Initial antigen immunization can be carried out using a Freund's adjuvant (CFA) and subsequent boosts can be carried out with an incomplete adjuvant. About 50-60 days after immunization, preferably on day 55, antibody titers are tested and the Antibody Selection (ABS) process begins if appropriate titers are established. The two key criteria for ABS initiation are recognition of potent antigens and function modifying activity in polyclonal serum.

At the time positive antibody titers are established, animals are euthanized and sources of B lymphocytes are isolated. These sources include: the spleen, lymph nodes, spinal cord and mononuclear cells of

478

peripheral blood (PBMC). Single cell suspensions are generated and the cell suspensions are washed to make them compatible for long term storage at low temperature. The cells are then typically frozen.

To start the antibody identification process, a small fraction of the frozen cell suspensions are thawed, washed, and placed in tissue culture medium.

These suspensions are then mixed with a biotinylated form of the antigen used to elicit the animal's immune response, and antigen-specific cells are recovered using Miltenyi's magnetic bead cell selection methods. Specific enrichment is carried out using streptavidin beads. The enriched population recovers and develops in the next phase of isolation of specific B lymphocytes.

Example 2 Production of clonal cultures containing antigen-specific B lymphocytes

The enriched B lymphocytes produced according to Example 1 are then plated at various cell densities per well in a 96-well microtiter plate. In general this is at 50, 100, 250 or 500 cells per well with 10 plates per group. The medium is supplemented with a conditioned medium of 4% rabbit activated T lymphocytes together with 50K of irradiated EL4B fibroblasts and

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FROZEN INDUSTRIAL PROPERTY MEXICAN INSTITUTE. These cultures remain unchanged for 5-7 days, at which time the secreted antibody containing supernatant is collected and evaluated for objective properties in a separate assay frame. The remaining supernatant is left intact and the plate is frozen at -70 ° C. Under these conditions, the culture process typically results in wells containing a mixed cell population comprising a clonal population of antigen-specific B lymphocytes, that is, a single well will only contain a single monoclonal antibody specific for the desired antigen.

Example 3 Analysis of antibody supernatants for monoclonal antibody of desired specificity and / or functional properties

Supernatants containing antibodies from the well containing an antigen-specific population of B lymphocytes produced according to Example 2, are initially analyzed for antigen recognition using ELISA methods. This includes selective immobilization of antigens (eg. , capture of biotinylated antigen by streptavidin-coated plates), plate coating for non-specific antigen or alternatively, through an antigen concentration strategy (eg, selective antigen capture followed by the addition of a binding to generate a heteromeric protein-antigen complex).

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INSTITUTO .MEXICANO CE LA 'RDPIEDAO industrial

Antigen-positive well supernatants are then optionally tested in a function-modification assay that is strictly ligand dependent. An example of these is an in vitro protein-protein interaction assay that recreates the natural interaction of the antigen ligand with the recombinant receptor protein. Alternatively, a ligand-dependent cell-based response is used and is easily monitored (eg, proliferation response). The supernatant showing significant antigen recognition and potency is considered a positive well. Cells from the original positive well then transition to the antibody recovery phase.

Example 4 Recovery of a Simple B Lymphocyte Producing Antibodies of Desired Antigen Specificity

Cells are isolated from a well containing a clonal population of antigen-specific B lymphocytes (produced according to Example 2 or 3) that secrete a single antibody sequence. The isolated cells are then tested to isolate a single cell that secretes antibodies. Dynal streptavidin beads are coated with biotinylated target antigens in buffered medium to prepare micro beads containing antigens compatible with cell viability. Then the antigen-loaded beads, cells that

481

they produce antibodies from a positive well and a fluorescein isothiocyanate-labeled anti-host IgG H&L antibody (FITC) (as noted, the host can be any mammalian host, e.g. rabbit, mouse, rat, etc. ) are incubated together at 37 ° C. This mixture is then re-pipetted into aliquots on a glass slide so that each aliquot has on average a single antibody-producing B lymphocyte. Antigen-specific and antibody-secreting cells are then detected by fluorescence microscopy. The secreted antibody is locally concentrated in adjacent beads due to bound antigen and provides location information based on the strong fluorescent signal. Antibody-secreting cells are identified by FITC detection of antibody-antigen complexes formed adjacent to the secretory cell. The single cell found in the center of this complex is then recovered using a micromanipulator. The cell is instantly frozen in an Eppendorf PCR tube for storage at -80 ° C until recovery of the antibody sequence begins.

Example 5 Isolation of Antibody Sequences from Antigen-Specific B Lymphocytes

Antibody sequences are recovered using a combined RT-PCR based method from a B lymphocyte.

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Mexican Institute of Industrial Property

single isolated according to Example 4 or an antigen-specific B-lymphocyte isolated from the clonal population of B lymphocytes obtained according to Example 2. Primers are designed to hybridize in conserved and constant regions of the target immunoglobulin genes (heavy and light), such as rabbit immunoglobulin sequences, and a two-step nested PCR recovery step is used. Amplicons from each well are analyzed to determine recovery integrity and size. The resulting fragments are then digested with AluI to genetically identify the cloning of the sequence. Identical sequences present a common fragmentation pattern in their electrophoretic analysis. Significantly, this common fragmentation pattern that tests cell cloning is generally observed even in wells originally plated up to 1000 cells / well. The original heavy and light chain amplicon fragments are digested by restriction enzyme with HindIII and Xhol or HindIII and BsiWI to prepare the respective pieces of DNA for cloning. The resulting digestions are then ligated into an expression vector and transformed into bacteria for the propagation and production of plasmids. Colonies are selected for sequence characterization.

Example 6 Recombinant production of monoclonal antibody of desired antigen specificity and / or functional properties

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

Correct full length antibody sequences are established for each well containing a single monoclonal antibody and a DNA miniprep is prepared using Qiagen solid phase methodology. DNA is then used to transfect mammalian cells to produce full-length recombinant antibodies. The crude antibody product is tested for antigen recognition and functional properties to confirm that the original characteristics are found in the recombinant antibody protein. When appropriate, transient mammalian transfections are completed on a large scale and the antibody is purified through Protein A affinity chromatography. Kd is evaluated using standard methods (eg Biacore ™) as well as IC50 in a potency assay.

Example 7 Preparation of Human IL-6 Binding Antibodies

Using the antibody selection protocol described herein, an extensive panel of antibodies can be generated. The antibodies possess high affinity for IL-6 (single and double digit pM Kd) and demonstrate potent IL-6 antagonism in multiple cell-based assay systems (T1165 and HepG2). Furthermore, the collection of antibodies presents different modes of antagonism towards the processes driven by IL-6.

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Immunization strategy

Rabbits were immunized with huIL-6 (R&R). Immunization consisted of a first subcutaneous injection (se) of 100 pg in a complete Freund's adjuvant (CFA) (Sigma) followed by two boosters, two weeks apart, of 50 pg each in incomplete Freund's adjuvant (IFA) (Sigma). Animals were bled on day 55 and serum titers were determined by ELISA (antigen recognition) and by non-radioactive proliferation assay (Promega) using the T1165 cell line.

Evaluation of the titration in the selection of antibodies

Antigen recognition was determined by coating Immulon 4 plates (Thermo) with 1 pg / ml huIL15 6 (50 pl / well) in phosphate buffered saline (PBS, Hyclone) overnight at 4 ° C. On the day of the assay, the plates were washed 3 times with PBS / Tween 20 (PBST tablets, Calbiochem). The plates were then blocked with 200 µl / well of 0.5% fish skin gelatin (FSG, Sigma) in PBS for 30 min at 37 ° C. The blocking solution was removed and the plates were transferred. Serum samples were made (bleed and prebleed) in an initial dilution of 1: 100 (all dilutions were made in 50 µg / well FSG) followed by 1:10 dilutions through the plate (column 12 was left in white for background control). Plates

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MEXICAN INSTITUTE OF THE INDUSTRIAL PROTÍEOAD

TO

A were incubated for 30 minutes at 37 ° C. The plates were washed 3 times with PBS / Tween 20. Goat anti-rabbit FC-HRP (Pierce) diluted 1: 5000 was added to all wells (50 µΙ / well), and the plates were incubated for 30 minutes at 37 ° C. The plates were washed as described above. 50 µΐ / well TMB-Stable (Fitzgerald Industries) was added to the plates and allowed to develop color, generally for 3 to 5 minutes. The growth reaction was stopped with 50 µΐ / well 0.5 M HC1. The plates were read at 450nm. Optical density (OD) against dilution was plotted using Graph Pad Prizm software and titrations were determined.

Assessment of functional qualification

The functional activity of the samples was determined by means of a T1165 proliferation assay. T1165 cells were routinely maintained in a modified RPMI medium (Hyclone) supplemented with Hepes, sodium pyruvate, sodium bicarbonate, L-glutamine, high glucose, penicillin / streptomycin, 10% heat inactivated fetal bovine serum (FBS ) (all Hyclone supplements), 2mercaptoethanol (Sigma) and 10 ng / ml huIL-6 (R&D) On the day of the test, cell viability was determined by trypan blue (Invitrogen) and cells were seeded at a fixed density of 20,000 cells / well. Before seeding, cells were washed twice in the medium described above without IL486

MEXJCA.NO INSTITUTE OF PROPERTY

INDUSTRIAL

human (spinning · at 13000 rpm for 5 minutes and discarding the supernatant). After the last wash, the cells were resuspended in the same medium used for washing in a volume equivalent to 50 µΐ / well. The cells were separated at room temperature.

In a 96-well round bottom plate (Costar), serum samples were added starting at 1: 100, followed by a 1:10 dilution through the plate (columns 2 to 10) at 30 µΐ / well in replicates of 5 (rows B to F: dilution carried out in the medium described above without huIL-6). The column only served as a medium for the control of IL-6. 30 µΐ / well of huIL-6 was added to all wells at a concentration of 4x final EC50 (previously determined concentration) (huIL-6 was diluted in the medium described above). The wells were incubated for 1 hour at 37 ° C to allow binding of the antibody to occur. After 1 hour, 50 µΐ / well of antibody-antigen complex (Ab-Ag) was transferred to a 96 well flat bottom plate (Costar) following the plate map format provided on the round bottom plate. In row G, 50 µΐ / well of medium was added to all wells (columns 2 to 11) for background control. 50 µΐ / well of the separated cell suspension was added to all wells (columns 2 to 11, rows B to G). In columns 1 and 12 and rows A and H, 200 μΐ / well of medium was added to prevent evaporation of

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test wells and to minimize edge effect. The plates were incubated for 72 h at 37 ° C in 4% CO2. At 72 hr, 20 µΐ / well of CellTiter96 reagents (Promega) were added to all test wells according to the manufacturer's protocol and the plates were incubated for 2 hr at 37 ° C. At 2 hr, the plates were gently mixed on an orbital shaker to disperse the cells and allow homogeneity in the test wells. The plates were read at a wavelength of 490 nm. Optical density (OD) versus dilution was plotted using Graph Pad Prizm software and functional titrations were determined. A positive assay control plate was made as described above using MAB2061 (R&D Systems) at an initial concentration of 1 pg / ml (final concentration) followed by 1: 3 dilutions through the plate.

Tissue culture

Once the acceptable titers were established, the rabbit (s) was slaughtered. The spleen, lymph nodes, and blood were cultured and processed as follows:

The spleen and lymph nodes were processed in a single-cell suspension by dissociating the tissue and opening a path using a sterile 70 μπι metal mesh (Fisher) with a plunger from a 20 cc syringe. The cells were collected in the described modified RPMI medium

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FROM THE INDUSTRIAL FROi-ISDAD previously without huIL-6 but with low glucose. The cells are. washed twice by centrifugation. After the last wash, the cell density was determined by trypan blue. Cells were spun at 1500 rpm for 10 minutes; supernatant was discarded. Cells were resuspended in the appropriate volume of 10% dimethyl sulfoxide (DMSO, Sigma) in FBS (Hyclone) and dispensed at 1 ml / container. The containers were then stored at -70 ° C for 24 h before being placed in a liquid nitrogen (LN2) tank for long-term storage.

Peripheral blood mononuclear cells (PBMC) were isolated by mixing blood with equal parts of the low glucose medium described above without FBS. 35 ml of the blood mixture was carefully layered over 8 ml of Lympholyte Rabbit (Cedarlane) in a 45 ml conical tube (Corning) and centrifuged for 30 minutes at 2500 rpm at room temperature without stopping. After centrifugation, the PBMC layers were carefully removed using a Pasteur glass pipette (VWR), combined and placed in a clean 50 ml container. Cells were washed twice with the modified medium described above by centrifugation at 1500 rpm for 10 minutes at room temperature and cell density was determined by trypan blue staining. After the last wash, cells were resuspended in an appropriate volume of 10% medium

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IMPI of DMSO / FBS and frozen as described above.

B lymphocyte culture

On the day the B cell culture was established, PBMC, spleen cells, or lymph nodes were thawed for use. The containers were removed from the LN2 tank and placed in a 37 ° C water bath until thawed. The contents of the containers were transferred to a 15 ml conical centrifuge tube (Corning) and 10 ml of modified RPMI described above was gently added to the tube. The cells were centrifuged for 5 minutes at 1.5K rpm and the supernatant was removed. Cells were resuspended in 10 ml of fresh medium. Cell density and viability were determined using trypan blue.

The cells were washed again and resuspended in 1E07 / 80 pL cells of medium. Biotinylated huIL-6 (B huIL-6) was added to the cell suspension at the final concentration of 3 pg / mL and incubated for 30 minutes at 4 ° C. Unbound B huIL-6 was removed with two 10 ml washes of buffered phosphate (PBF): Ca / PBS Mg-free (Hyclone), 2 mM ethylenediamine tetraacetic acid (EDTA), 0.5% bovine serum albumin (BSA ) (Sigma-free biotin). After the second wash, the cells were resuspended in 1E07 / 80 pl cells of PBF. Added 20 pl of streptavidin MACS® beads

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY (Milteni) / 10E7 cells to the cell suspension. Cells were incubated at 4 ° C for 15 minutes. Cells were washed once with 2 ml of PBF / 10E7 cells. After washing, the cells were resuspended in 1E08 / 500 µΐ PBF cells and separated. A MACS® MS column (Milteni) was prewashed with 500 ml of PBF in a magnetic mechano (Milteni). The cell suspension was applied to the column through a pre-filter and an unbound fraction was collected. The column was washed with 1.5 ml of PBF buffer. The column was removed from the magnetic mechano and placed in a clean 5 ml sterile Falcon-type polypropylene tube. 1 ml of PBF buffer was added to the top of the column and positive selected cells were collected. The yield and viability of the positive and negative cell fractions were determined by trypan blue staining. Positive selection provided an average of 1% of the initial cell concentration.

A pilot cell analysis was established to provide information about the seeding levels for the culture. Three groups of 10 plates (a total of 30 plates) were seeded at 50, 100 and 200 enriched B lymphocytes / well.

Additionally, each well contained 50K cells / well of irradiated EL-4.B5 cells (5,000 Rads) and an appropriate level of T cell supernatant (ranging from 1-5% depending on preparation) in a modified high glucose medium RPMI at a final volume of 250 μΐ / well. The cultures were incubated

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for 5 to 7 days at 37 ° C in 4% CO2

Identification of B lymphocytes that secrete selective antibodies

<td>The</td><td>crops were tested among the</td><td>days</td><td> 5</td><td>and 7 for</td>

<td>decide</td><td>antigen recognition</td><td>and</td><td>the</td><td>exercise</td>

<td>functional.</td><td></td><td></td><td></td><td></td>

<td colspan="2">Antigen recognition analysis</td><td></td><td></td><td></td>

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above, except that 50 µΐ of supernatant from B-cell culture wells (BCC) (all 30 plates) was used as the source of the antibody. The conditioned medium was transferred to antigen-coated plates. After positive plates were identified, the supernatant was removed and transferred to one or more 96-well main plates. Then the original culture plates were frozen removing all supernatants except 40 µΐ / well and adding 60 µΐ / well of 16% DMSO in FBS. The plates were wrapped in paper towels to slow down the freezing and placed on

-70 ° C.

Functional activity analysis

Main plates were then analyzed for functional activity in the proliferation assay

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T1165 as described above, except for row B which was medium only for background control, row C which was medium + IL-6 for positive control of proliferation and rows DG and columns 2-11 which were the wells of BCC (50 μΐ / well, single points). 40 µL of IL-6 was added to all wells except the middle row at 2.5 times the EC50 concentration determined for the assay. After 1 hr incubation, the Ab / Ag complex was transferred to a 96-well flat-bottom plate treated with tissue culture (CT). 20 µΐ of cell suspension in a modified RPMI medium without huIL-6 (T1165 at 20,000 cells / well) (100 µ / final volume per well) was added to all wells. The background was subtracted and the observed OD values were transformed into% inhibition.

B lymphocyte recovery

Plates containing wells of interest were removed at -70 ° C and cells from each well were recovered with 5-200 µΐ medium / well washes. The washes were accumulated in a sterile 1.5 ml centrifuge tube and the cells were pelleted for 2 minutes at 1500 rpm.

The tube was inverted, the spin was repeated, and the supernatant was carefully removed. The cells were resuspended in 100 µΐ / tube of medium. 100 μΐ of streptavidin M280 biotinylated dynabeads were added to the cell suspension

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coated with IL-6 (Invitrogen) and 16 µΐ goat anti-rabbit IgG-FITC H&L diluted 1: 100 in the medium.

20 µΐ were removed from a cell / beads / FITC suspension and 5 µΐ droplets were prepared on a glass slide (Corning) previously treated with

Sigmacote (Sigma), 35 to 40 droplets / slide. A waterproof kerosene barrier (JT Baker) was added to immerse the droplets and the slide was incubated for 90 minutes at 37 ° C with 4% CO2 in the dark.

Antibody-producing specific B lymphocytes can be identified by the fluorescent ring around them due to antibody secretion, recognition of the biotinylated antigen associated with beads, and subsequent detection by the IgG fluorescent detection reagent. Once a cell of interest was identified, the cell in the center of the fluorescent ring was recovered through a micromanipulator (Eppendorf). The only cell that synthesized and exported the antibody was transferred to a 250 pL microcentrifuge tube and placed on dry ice. After recovering all the cells of interest, they were transferred to -70 ° C for long-term storage.

Example 8 Expression of yeast cells

Antibody genes: Genes were cloned and interpreted to direct the synthesis of an antibody

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INSTITUTO MEXICANO monoclonal rabbit humanized chimeric. OFIndustrial

Expression vector: The vector contains the functional component signs: 1) a mutant origin of replication ColEl, which facilitates the replication of the plasmid vector in cells of the bacterium Escherichia coli, 2) a bacterial gene

Sh ble that confers resistance to the antibiotic Zeocin ™ (phleomycin) and serves as a selectable marker for transformations of both E. coli and P. shepherds, 3) an expression cassette composed of the promoter of the glyceraldehyde dehydrogenase gene (GAP gene) fused to the sequences that encode the leader sequence of secretion pre pro of the alpha mating factor of Saccharomyces cerevisiae, followed by sequences that encode a signal transcriptional termination of the P. pastoras alcohol oxidase I gene (AOX1). The Zeocin ™ resistance marker gene (phleomycin) provides an enrichment medium for strains containing multiple integrated copies of an expression vector in a strain by selecting transformers resistant to high levels of Zeocin ™ (phleomycin).

P. pastoras strains: You can use the P. pastoras strains metí, lys3, ura3 and adel. Although any two complementary sets of auxotrophic strains can be used for the construction and maintenance of diploid strains, these two strains are especially suitable for this method for two reasons. First, they grow slower than diploid strains

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which are the result of their mating or merging. Therefore, if a small number of adel or ura3 haploid cells are still present in a culture or arise through meiosis or other mechanism, the diploid strain should grow further in the culture.

Second, it is easy to monitor the sexual status of these strains, since the Ade + diploid colonies that arise from their mating are white or regular cream, while the cells of any strain that are adelo haploid mutants will form a colony with a pink color. distinctive. Furthermore, any strains that are ura3 haploid mutants are resistant to 5-fluoro-orotic acid (FOA) drug and can be sensibly identified by plating samples of a culture in minimal medium + uracil plates with FOA. In these plates, only uracil mutant strains that require uracil (supposedly haploid) can grow and form colonies. Thus, with adel and ura3-labeled haploid progenitor strains, the sexual status of the resulting antibody-producing diploid strains (haploid versus diploid) can be easily controlled.

Methods

Construction of pGAPZ-alpha expression vectors for transcription of light and heavy chain antibody genes. Humanized heavy and light chain fragments were cloned into pGAPZ expression vectors through a process

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY directed to PCR. The recovered humanized constructs were subjected to amplification under conditions of a standard KOD polymerase kit (Novagen) ((1) 94 ° C, 2 minutes; (2) 94 ° C, 30 · seconds (3) 55 ° C, 30 seconds ; (4) 72 ° C, 30 seconds, alternated through steps 2-4 35 times; (5) 72 ° C 2 minutes) using the following primers (1)

AGCGCTTATTCCGCTATCCAGATGACCCAGTC direct light chain-the Afel site has simple underlining. The end of the HSA signal sequence is underlined twice, followed by the mature variable light chain sequence (not underlined); the

CGTACGTTTGATTTCCACCTTG inverse.

Variable Light Chain Reverse Primer. The BsiWI site is underlined, followed by the reverse complement for the 3 'end of the variable light chain. Digestion of the restriction enzyme with Afel and BsiWI allows in-frame insertion with the vector pGAPZ using the human HAS leader sequence in-frame with the human kappa light chain constant region for export. (2) A similar strategy is carried out for the heavy chain. The direct primer used is AGCGCTTATTCCGAGGTGCAGCTGGTGGAGTC. The Afel site has simple underlining. The end of the HSA signal sequence is underlined twice, followed by the sequence for the mature variable heavy chain (not underlined). The reverse heavy chain primer is CTCGAGACGGTGACGAGGGT. The Xhol site is underlined, followed by the reverse complement for the 3 'end of the heavy chain

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variable. This allows cloning of the heavy chain in-frame with the IgG-Ql CH1-CH2-CH3 region previously inserted into pGAPZ using a comparable directional cloning strategy.

Transformation of expression vectors in haploid host strains adel ura3, metí and lys3 of P. pastoris. All methods used for transformation of haploid strains of P. pastoris and genetic manipulation of the P. pastoris sexual cycle are described in Higgins, DR, and Cregg, JM, Eds. 1998. Pichia Protocols. Methods in Molecular

Biology. Humana Press, Totowa, NJ.

Prior to transformation, each expression vector is linearized within the GAP promoter sequences with Avrll to direct the integration of the vectors into the GAP promoter locus of the P. pastoris genome. Samples from each vector are then individually transformed into electrocompetent cultures of adel, ura3, meti and lys3 strains by electroporation and successful transformers on YPD Zeocin ™ plates (phleomycin) are selected for their resistance to this antibiotic. The resulting colonies are selected, scored for single colonies on YPD Zeocin ™ (phleomycin) plates, and then examined for the presence of an antibody gene insert by PCR assay on genomic DNA extracted from each strain for the insert of suitable antibody gene and / or by the capacity of each strain of

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synthesize an antibody chain by a colony blot / blot method (Wung et al. Biotechniques 21 808-812 (1996). Haploid adel, meth and lys3 strains expressing one of three heavy chain constructs are harvested for constructs diploid along with a haploid ura3 strain that expresses a light chain gene. The heavy chain genes expressing haploids are paired with the appropriate light chain haploid ura3 to generate a diploid secretory protein.

Pairing of haploid strains that synthesize a single antibody chain and selection of diploid derivatives that synthesize tetrameric functional antibodies. To pair the haploid strains of P. pastoris, each strain that produces an adel (or meth or lys3) heavy chain that is to be crossed was streaked through a YPD-rich plate and the strain that produces an ura3 light chain was streaked to through a second plate

YPD (~ 10 grooves per plate). After a day or two of incubation at 30 ° C, cells from a plate containing heavy chain strains and a plate containing ura3 light chain strains are transferred to a sterile velvet cloth on a replica plate block in a incubation cross standard so that each heavy chain strain contains a patch of cells mixed with each light chain strain. Cells plated on cross and streak replica plates are then transferred to a mating plate and incubated at 25 ° C

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY to stimulate the initiation of mating between the strains. After two days, cells in the mating plates are transferred back to sterile velvet on a replica plate block and then transferred to minimal media plates. These plates are incubated at 30 ° C for three days to allow selective growth of colonies of prototrophic diploid strains. The colonies that emerged are chosen and streaked onto a second plate of minimal medium to isolate the single colony and purify each diploid strain. The resulting diploid cell lines are then examined to determine antibody production.

The putative diploid strains are tested to demonstrate that they are diploid and that they contain both expression vectors for antibody production. For diploidy, samples of a strain are propagated on mating plates to stimulate them to undergo meiosis and form spores. Haploid spore products are collected and tested for phenotype. If a significant percentage of the resulting spore products are single or double auxotrophs, it can be concluded that the original strain must have been diploid. Diploid strains are examined for the presence of both antibody genes by extracting the genomic DNA from each and using this DNA in PCR reactions specific to each gene.

Fusion of haploid strains that synthesize a chain of a

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INDUSTRIAL single antibody and selection of diploid derivatives that synthesize tetrameric functional antibodies. As an alternative to the pairing procedure described above, single-chain antibody single cultures producing haploid strains of adel and ura3 are spheroplastized and their resulting spheroplasts are fused using polyethylene glycol / CaCl2. The fused haploid strains are then placed on agar containing 1M sorbitol and minimal medium to allow the diploid strains to regenerate their cell wall and develop into visible colonies. The resulting colonies are collected from the agar and streaked on a minimal medium plate and the plates are incubated for two days at 30 ° C to generate colonies from single cells from diploid cell lines. The putative resulting diploid cell lines are then examined to determine diploidy and antibody production as previously described.

Antibody purification and analysis. A diploid strain for full length antibody production is derived through pairing of the meth light chain and the lys3 heavy chain using the methods described above. Culture media from cultures of the P. pastoris diploid expression strain shaker or fermenter flask are collected and examined for the presence of the antibody protein via SDS-PAGE e

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heavy and light chains of human IgG or specifically against the IgG heavy chain.

To purify yeast secreted antibodies, the clarified antibody-producing culture media is passed through a Protein A column and after washing with 20 mM sodium phosphate pH 7.0 and binding buffer, the protein bound to the Protein A is eluted using 0.1 M glycine HC1 buffer, pH 3.0. Fractions containing most of the total protein are examined by Coomasie blue stained SDS-PAGE and immunoblotting for antibody proteins. The antibody is characterized using the ELISA described above for IL-6 recognition.

Antibody activity assay. The yeast-derived recombinant antibody is evaluated for functional activity through the IL-6 driven T1165 cell proliferation assay and the IL-6 stimulated HepG2 haptoglobin assay described above.

Example 9 Neutralization of the acute phase response by intravenous administration of anti-IL-6 Abl antibody.

Human IL-6 can elicit an acute phase response in rats and one of the most important acute phase proteins that is stimulated in the rat is α-2 macroglobulin (A2M). A study was designed to assess Abl antibody dose

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required to eradicate the A2M response to a single 100 pg sc injection of human IL-6 administered one hour after the different doses (0.03, 0.1, 0.3, 1 and 3 mg / kg) of the Abl antibody administered intravenously ( n = 10 rats / dose level) or polyclonal human IgGl as control (n = 10 rats). Plasma was recovered and A2M was quantified using a commercial ELISA kit (ICL Inc.,

Newberg OR; cat. no.- E-25A2M). The endpoint was the difference in plasma A2M concentration at the 24 hour time point (after Abl). The results are shown in Figure 4.

The ID50 for the Abl antibody was 0.1 mg / kg with complete suppression of the A2M response at 0.3 mg / kg. This firmly establishes that in vivo neutralization of human IL-6 can be accomplished by the Abl antibody.

Example 10 RXF393 Cachexia Model Study 1

Introduction

The human kidney cancer cell line, RXF393, produces significant weight loss when transplanted into nude hairless mice. Weight loss begins around day 15 after transplantation and 80% of animals lose at least 30% of their total body weight around days 18-20 after transplantation. RXF393 secretes human IL-6 and the plasma concentration of human IL-6 in these animals is very

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TIUAL iNDl high around 10 ng / ml. Human IL-6 can bind the murine soluble IL-6 receptor and activate mouse IL6 responses. Human IL-6 is approximately 10 times less potent than murine IL-6 in activating responses of

IL-6 in the mouse. The objectives of this study were to determine the effect of the Abl antibody on survival parameters, body weight, serum amyloid protein A, hematology and tumor growth in nude athymic mice transplanted with the human kidney cancer cell line

RXF393.

Methods

Eighty 6-week-old male nude athymic mice had RXF393 tumor fragments (30-40 mg) implanted subcutaneously on the right side. The animals were then divided into eight groups of ten mice. Three groups were given 3 mg / kg, 10 mg / kg or 30 mg / kg of Abl antibody intravenously weekly on day 1, day 8, day 15 and day 22 after transplantation (evolution groups). Three other groups were given 3 mg / kg, 10 mg / kg or 30 mg / kg of Abl antibody intravenously weekly on day 8, day 15 and day 22 after transplantation (regression groups). Finally, a control group was given 30 mg / kg of polyclonal human IgG and a second control group was given phosphate buffered saline intravenously weekly on day 1, day

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8, day 15 and day 22 after the transplant. Industrial - Animals euthanized ul dícrT ^ ST ^ uañ the tumor reached 4,000 mm3 or when they weakened (> 30% loss of body weight). Animals were weighed on days 1 and 6 and then daily from days 9 to 28 after transplantation. Average body weight percentage (MPBW) was used as the main parameter to control weight loss during the study. The following was calculated: (body weight-tumor weight) / reference body weight x 100. Tumor weight was measured on days 1, 6, 9, 12, 15, 18, 22, 25 and 28 after transplant. Under anesthesia, blood was drawn from five mice in each group on days 5 and 13 and from ten mice in each group when euthanized (in most cases on day 28). Blood was analyzed to determine hematology and serum amyloid and protein A (SAA) concentration. Blood samples were drawn from an additional group of 10 6-week-old athymic nude mice without tumor to estimate hematology and SAA concentration as a set of reference values.

Results - Survival

No animals were euthanized or killed in any Abl antibody group prior to the study end date on day 28. In the two control groups, 15 animals (7/9 in the polyclonal human IgG group)

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and 8/10 in the phosphate buffered saline group) were found dead or euthanized because they were severely weakened (> 30% loss of body weight). The median survival time in both control groups was days.

Survival curves for the two control groups and the Abl antibody evolution groups (dosed from day 1 of the study) are presented in

Figure 5.

Survival curves for the two control groups and the Abl antibody regression groups (dosed from day 8 of the study) are presented in Figure 6.

There was a statistically significant difference between the survival curves of the polyclonal human IgG (p = 0.0038) and phosphate buffered saline (p = 0.0003) control groups and the survival curve of the six Abl antibody groups. There was no statistically significant difference between the two control groups (p = 0.97).

Results - Tumor size

The size of the tumor in the surviving mice was estimated by palpation. In the first 15 days of the study, none of the mice in either group were found dead or euthanized and therefore the size comparison

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Tumor IMPI between groups these days was free from sampling biases. No difference in tumor size was observed between the evolution or regression groups of the Abl antibody and the control groups until day 15. A comparison of tumor sizes between surviving mice in the control and treatment groups was not performed after the onset of mortality in the controls (on day 15), since the tumor size in surviving control mice was allegedly biased. and therefore the results of that comparison would not be significant.

Because administration of the Abl antibody promoted survival without apparent reduction in tumor size, elevated serum IL-6 may contribute to mortality by independent tumor growth mechanisms. These observations support the hypothesis that the Abl antibody may promote cancer patient survival without directly affecting tumor growth, possibly enhancing the well-being of the patient as a whole.

Results - Weight loss

The average body weight percentage (MPBW) (□ SEM) against time is shown in Figure 27. Compared to controls, Abl dosed mice were protected against weight loss. On day 18, the MPBW in control mice was 75%, corresponding to a loss of

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INDUSTRIAL average weight of 25%. In contrast, on the same day, the MPBW in Ab-1 treatment groups had changed minimally (between 97% and 103%). There was a statistically significant difference between the MPBW curves in controls (receiving polyclonal human IgG or PBS) and those in the 10 mg / kg dosage group (p <0.0001) or the 3 mg / kg and 30 dosage groups mg / kg (p <0.0005). There was no statistically significant difference between the two control groups.

Representative photographs of control and Abl-treated mice (Figure 28) illustrate the emaciated condition of control mice compared to the normal appearance of the Abl-treated mouse at the end of the study (note externally visible tumor sites on the side straight).

These results suggest that Abl may be useful in preventing or treating cachexia caused by elevated IL-6 in humans.

Results - Serum amyloid plasma A

The mean serum amyloid plasma A concentration (± SEM) against time in the two control groups and the evolution (dosed from day 1 of the study) and regression (dosed from day 8 of the study) groups of the Abl antibody They are presented in Table 5 and graphically in Figure 32.

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Table 5: Average plasma SAA-Abl antibody, all groups against control groups

<td rowspan="3"></td><td colspan="3">Plasma</td><td colspan="3" rowspan="2">Plasma average</td><td colspan="2" rowspan="3">Plasma average SAA ± SEM bleeding terminal (pg / ml)</td>

<td colspan="2" rowspan="2">average SAAiSEM (pg / ml)</td><td rowspan="2">Day 5</td>

<td>polyclonal iv</td><td>(n = 5)</td><td></td><td></td><td>(n = 4)</td><td></td><td></td><td>(n = 4)</td><td></td>

<td>weekly</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>weekly</td><td>(n = 5)</td><td></td><td></td><td>(n = 5)</td><td></td><td></td><td>(n = 3)</td><td></td>

<td>weekly</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>weekly</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

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<td rowspan="2">Abl 3 mg / kg iv weekly from day 1</td><td colspan="3" rowspan="2">106 ± 9 (n = 5)</td><td rowspan="2">1623 (n = 4)</td><td rowspan="2"> +</td><td colspan="3"></td>

<td>weekly</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>weekly</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td>weekly</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

SAA is up-regulated through hIL-6 stimulation and this response is directly correlated with circulating levels of hIL-6 arising from the implanted tumor. The surrogate marker 5 provides an indirect reading of active hIL-6. Therefore, in the two treatment groups described above, there are significantly decreased levels of SAA due to the neutralization of hIL-6 that comes from a tumor. It further supports the discussion that the

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Abl antibody shows efficacy in vivo.

Example 11 Study 2 of the cachexia model RXF393

Introduction

A second study was conducted in the RXF-393 cachexia model where treatment with the Abl antibody began at a later stage (days 10 and 13 after transplantation) and with a longer treatment phase (up to 49 days after transplantation). . The dosage interval with the Abl antibody was shortened from 7 to 3 days and the daily food consumption was also measured. There was also an attempt to standardize tumor sizes at the time of dosing with the Abl antibody.

Methods

Eighty 6-week-old male nude athymic mice had RXF393 tumor fragments (30-40 mg) implanted subcutaneously on the right side. Twenty mice whose tumors had grown to between 270-320 mg in size were selected and divided into two groups. One group received the Abl antibody at 10 mg / kg iv every three days and the other group received 10 mg / kg of polyclonal human IgG every 3 days thereafter (day 10 after transplantation). Other mice were selected when their tumor size had reached 400-527 mg in size and divided into two groups. One group received the

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Abl antibody at 10 mg / kg iv every three days and eT ^ other

moment (day 13 after transplant). The remaining 40 mice were no longer part of the study and were euthanized on day 49, when the tumor reached 4,000 mm.3 or when they became very weak (> 30% loss of body weight).

Animals were weighed every 3-4 days from day 1 to day 49 after transplantation. Average body weight percentage (MPBW) was used as the main parameter to control weight loss during the study. It was calculated in this way: ((body weight - tumor weight) / reference body weight) x 100. Tumor weight was measured every 3-4 days from day 5 to day 49 after transplantation. Food consumption was measured each day (amount consumed in hours in weight (g) per treatment group) from day 10 in the 270-320 mg tumor groups and from day 13 in the 400 tumor groups -527 mg.

Results - Survival

Survival curves for Abl antibody at 10 mg / kg iv every three days (tumor size 270-320 mg) and 10 mg / kg iv for polyclonal human IgG iv every three days (tumor size 270- 320 mg) are presented in Figure 7.

Median survival for Abl antibody at 10

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mg / kg iv every three days (tumor size 270-320 mg) was

............ 1 1 ·β^^^ ““ · ^ β * · »^^ βιιι ·· * ιιι ™ ιι ·· I IR of 46 days and 10 mg / kg iv for polyclonal human IgG every three days (tumor size 270-320 mg) was 32.5 days (p = 0.0071).

Survival curves for Abl antibody at 10 mg / kg iv every three days (tumor size 400-527 mg) and at 10 mg / kg iv for polyclonal human IgG every three days (tumor size 400-527 mg ) are presented in Figure 8. Median survival for Abl antibody at 10 mg / kg iv every three days (400-527 mg tumor size) was 46.5 days and at 10 mg / kg iv for polyclonal human IgG every three days (tumor size 400-527 mg) was 27 days (p = 0.0481).

Example 12 Multiple dose pharmacokinetic evaluation of Abl antibody in non-human primates

The Abl antibody was dosed in a single bolus infusion to a single male and a single female macaque monkey in phosphate buffered saline. Plasma samples were removed at fixed time intervals and the Abl antibody level was quantified through the use of an antigen capture ELISA assay. Biotinylated IL-6 (50 µΐ 3 pg / mL) was captured in streptavidin-coated 96-well microtiter plates. The plates were washed and blocked with 0.5% fish skin gelatin. The

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Properly diluted plasma samples were added and incubated for 1 hour at room temperature. Supernatants were removed and a second conjugated anti-hFc-HRP antibody was applied and left at room temperature.

The plates were then aspirated and TMB added to visualize the amount of antibody. Specific levels were then determined using a standard curve. A second dose of Abl antibody was administered on day 35 to the same two macaque monkeys and the experiment was replicated using an identical sampling plan. The resulting concentrations are then plotted against time, as shown in Figure 9.

This expressed and purified full-length humanized aglycosylated antibody Pichia pastoris shows characteristics comparable to the expressed mammalian protein. Furthermore, multiple doses of this product show reproducible half-lives, inferring that this production platform does not generate products that show enhanced immunogenicity.

Example 13 Mechanical characterization of antibody proteins with Octet

IL-6 signaling depends on the interactions between IL-6 and two receptors, IL-6R1 (CD126) and gpl30 (IL-6 signal transducer). To determine the mechanism of action of the antibody, mechanical studies were performed using biofilm interferometry with an Octet QK instrument.

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(ForteBio; Menlo Park, CA). Studies were conducted in two different configurations. In this first orientation, biotinylated IL6 (R&D system part number 206-IL-001MG / CF, biotinylated using Pierce EZ-link sulfo-NHS-LC-LC-biotin, product number 21338 according to manufacturer's protocols ) was initially linked to a streptavidin-coated biosensor (part number ForteBiol8-5006). Bonding is controlled as a signal boost.

The IL-6 bound to the sensor was then incubated with the antibody in question or with the diluent solution alone. The sensor was then incubated with a soluble IL-6R1 molecule (R&D systems product number 227-SR-025 / CF). If the IL6R1 molecule was unable to bind, the antibody was understood to block IL-6 / IL-6R1 interactions. These complexes were incubated with gpl30 (R&D systems 228-GP-010 / CF) in the presence of IL-6R1 for stability purposes. If gpl30 did not bind, it was concluded that the antibody blocked gpl30 interactions with IL-6.

In the second orientation, the antibody was bound to a biosensor coated with an anti-human Fe IgGl specific reagent (part number ForteBiol8-5001). IL-6 bound to the immobilized antibody and the sensor was incubated with IL-6R1. If IL-6R1 did not interact with IL-6, then it was concluded that the IL-6 binding antibody blocked IL6 / IL-6R1 interactions. In situations where antibody / IL6 / IL-6R1 is observed, the complex was incubated with gp! 30 in the presence of IL515

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6R1. If the gpl30 did not interact, it was then concluded that the antibody blocked IL-6 / gpl30 interactions. All studies were performed in a final volume of 200 pL at 30 ° C and 1000 rpm. For these studies, all proteins were diluted using a sample of ForteBio diluent buffer (part number 18-5028).

The results are presented in Fig. 10 (AE) and in the

Fig. 11.

Example 14 Peptide mapping

To determine the epitope recognized by Abl in human IL-6, the antibody was used in a Western blot-based assay. The human IL-6 form used in this example was 183 amino acids long (shown below). A 57-member library of overlapping 15 amino acid peptides comprising this sequence was commercially synthesized and covalently linked to a PepSpots nitrocellulose membrane (JPT Peptide technologies, Berlin, Germany). The overlapping 15 amino acid peptide sequences are shown in Figure 12. The bands were prepared and probed according to manufacturer's recommendations.

Briefly, the bands were pre-soaked in methanol, rinsed with PBS and blocked for more than 2 hours in 10% skim milk in PBS / 0.05% Tween (blocking solution). The

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INDUSTRIAL '«« the Abl antibody was used in a final dilution of 1 mg / ml and the secondary HRP-conjugated mouse Anti-Human-Kappa antibody (Southern BioTech # 9220-05) was used in a dilution of 1: 5000.

Antibody dilutions / incubations were done in a blocking solution. Transfers were performed using documented Amersham ECL reagents (GE # RPN2135) and chemiluminescent signal using a CCD camera (Alphalnnotec). The results of the transfers are shown in Figures 13 and 14.

The sequence of the human IL-6 form used to generate a library of peptides is set out below:

VPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENNLNL

PKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQ

KKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKEFLQSSLRALRQM (SEQ

ID NO: 1).

Example 15 Abl has high affinity for IL-6

A surface plasmon resonance was used to measure the association rate (Kto), dissociation rate (Kd) and dissociation constant (KD) for Abl for rat, mouse, dog, human and macaque monkey IL-6 at 25 ° C (Figure 15A). The dissociation constant of human IL-6 was 4 pM, indicating a very high affinity. As expected, the affinity generally decreased with the human's phylogenetic distance. The dissociation constants of Abl for IL-6 of macaque monkeys,

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respectively. Abl's affinity for iL-b ae was below the quantification limit of the experiment.

The high affinity of Abl for mouse, rat and macaque monkey IL-6 suggests that Abl can be used to inhibit IL-6 from these species. This hypothesis was tested using a cell proliferation assay. In summary, each IL-6 from these species was used to stimulate the proliferation of T1165 cells and the concentration at which Abl could inhibit 50% of the proliferation (IC50) was measured. The inhibition coincided with the measured dissociation constants (Figure 15B). These results demonstrate that Abl can inhibit native IL-6 of these species and suggest the use of these organisms to perform in vitro or in vivo models of inhibition of IL-6 by Abl.

Example 16 Multiple dose pharmacokinetic evaluation of the Abl antibody in healthy human volunteers.

The Abl antibody was dosed to healthy human volunteers in a single bolus infusion of histidine and sorbitol. The 1 mg, 3 mg, 10 mg, 30 mg or 100 mg doses were administered to each individual in dosage groups containing five to six individuals. Plasma samples were removed at fixed time intervals for up to twelve weeks. Human plasma was collected through venipuncture into a tube.

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INDUSTRIAL vacuum collection containing EDTA. Plasma was separated and used to assess Abl circulation levels using an Abl-specific monoclonal antibody, as follows. A 96-well microtiter plate was coated overnight with Abl-specific monoclonal antibody in IX

PBS overnight at 4 ° C. The remaining steps were carried out at room temperature. The wells were aspirated and subsequently blocked using 0.5% fish skin gelatin (FSG) (Sigma) in PBS IX for 60 minutes. Human plasma samples were then added and incubated for 60 minutes, then aspirated, then 50 pL of 1 pg / mL biotinylated IL-6 was added to each well and incubated for 60 minutes. The wells were aspirated and 50 pL streptavidin-HRP (Pharmingen) diluted 1: 5,000 in 0.5% FSG / PBS were added and incubated for 45 minutes. Development was done using standard methods using TMB for detection. The levels were then determined by comparison with a standard curve prepared in a comparable format.

The average plasma Abl concentration for each dosing group versus time is shown in Figure 16. AUC and C averagesmax they increased linearly with the dosage (Figures 17 and 18, respectively). For doses of 30 mg and more, the average Abl half-life in each dosing group was approximately 25-30 days (Figure

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cancer patients a single infusion of phosphate into five individual received a

Ex murmur 17 Abl Pharmacokinetics in advanced

The Abl antibody was dosed bolus into buffered saline individuals with advanced cancer. Each dose of 80 mg (n = 2) or 160 mg (n = 3) of Abl. Plasma samples were drawn weekly and the Abl antibody level was quantitated as in Example 16.

The average plasma Abl concentration in these individuals as a function of time is shown in Figure 20. The average Abl half-life was approximately 31 days.

Ex & aplo IB Half-life Abl unprecedented

In general, the average Abl half-life was approximately 31 days in humans (for dosages of 10 mg and more) and approximately 15-21 days in macaque monkeys. The half-life of Abl in humans and macaque monkeys is unprecedented when compared to the half-life of other anti-IL6 antibodies (Figure 21). As described above, Abl came from the humanization of a rabbit antibody and is produced from Pichia pastoris in aglycosylated form. These characteristics result in an antibody with very little immunogenicity in humans. Furthermore, the lack of glycosylation

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MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY prevents Abl from interacting with the Fe receptor or complement. Without wishing to be bound by theory, Abl's unprecedented half-life is believed to be at least partially attributable to humanization and lack of glycosylation. The particular sequence and / or structure of antigen binding surfaces may also contribute to the half-life of

Abl.

Example 19 Effect of Abl on Hemoglobin Concentration, Plasma Lipid Concentration, and Neutrophil Counts in Patients with Advanced Cancer

The Abl antibody was dosed in a single bolus infusion in phosphate buffered saline to eight individuals with advanced cancer (NSCLC, colorectal cancer, cholangiocarcinoma, or mesothelioma). Each individual received a dose of 80 mg, 160 mg, or 320 mg of Abl. Blood samples were drawn just prior to infusion and at fixed time intervals for six weeks, and hemoglobin concentration, plasma lipid concentration, and neutrophil counts were determined. The average hemoglobin concentration increased slightly (Figure 22), as did total cholesterol and triglycerides (Figure 23), while the average neutrophil counts decreased slightly (Figure 24).

These results further demonstrate some of the

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beneficial effects of the administration of Abl to individuals with chronic diseases. Because IL-6 is the major cytokine responsible for anemia in chronic diseases (including cancer-related anemia), neutralization of IL-6 by Abl increases the concentration of hemoglobin in these individuals. Similarly, because IL-6 is very important in increasing neutrophil counts in inflammation, the slight reduction seen in neutrophil counts further confirms that Abl inhibits IL-6. Finally, IL-6 causes anorexia as well as cachexia in these patients; neutralization of IL-6 by Abl causes appetite to recover and cachexia to be reversed. The increase in plasma lipid concentrations reflects the improved nutritional status of the patients. Taken together, these results further demonstrate that Abl effectively reverses these adverse consequences of IL6 in these patients.

Example 20 Abl suppresses serum CRP in healthy volunteers and in patients with advanced cancer

Introduction

Serum CRP concentrations have been described as a strong prognostic indicator in patients with some forms of cancer. For example, Hashimoto et al. conducted univariate and multivariate analyzes of CRP concentrations in

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103 (9): 1856-1864 (2005)). Patients were classified into two groups, those with serum CRP levels> 1.0 mg / dL (group with positive CRP) and those with serum CRP levels <1.0 mg / dL (group with negative CRP). The authors described an important correlation between preoperative serum CRP level and tumor size. Id

Additionally, the authors found that [the] overall survival and recurrence-free survival rates in the group with positive CRP were significantly lower compared to the rates in the group with negative CRP. Id. The authors concluded that the preoperative CRP level in patients is an important and independent predictive indicator of poor prognosis and early recurrence in patients with hepatocellular carcinoma.

Other researchers have identified similar correlations. For example, Karakiewicz et al. determined that serum CRP was an independent and informative indicator of the specific mortality of renal cell carcinoma (Karakiewicz, PI, et al., Cancer, 110 (6): 1241-1247 (2007)).

Accordingly, there remains a need in the art for methods and / or treatments that reduce serum C-reactive protein (CRP) concentrations in cancer patients and,

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MEXICAN INSTITUTE particularly, in those with advanced cancers ^ ü ^ B-wm

Methods

Healthy volunteers received a single 1-hour intravenous (IV) infusion of 100 mg (5 patients), 30 mg (5 patients), 10 mg (6 patients), 3 mg (6 patients), or 1 mg (6 patients) of the Abl monoclonal antibody, while 14 other healthy volunteers received intravenous placebo. Comparatively, 2 patients with advanced forms of colorectal cancer received a single 1-hour intravenous (IV) infusion of 80 mg of the monoclonal antibody Abl. No additional doses of the Abl monoclonal antibody were administered to the test population.

Patients were evaluated prior to dose administration and thereafter weekly for at least 5 weeks after dose. At the time of each evaluation, the serum CRP concentration in the patients was analyzed.

Results

Healthy volunteers

As mentioned above, serum CRP levels are a marker of inflammation, therefore, reference CRP levels are typically low in healthy individuals. Low reference CRP levels can make it difficult to detect a further reduction in CRP levels. However, a reduction was detected

524

Considerable IMPI of serum CRP concentrations in healthy volunteers who received all concentrations of the monoclonal antibody Abl, compared to controls (Figure 25). The reduction in serum CRP levels was rapid, occurred within one week from the administration of the antibody, and lasted at least until the final measurement was made (8 or 12 weeks from the administration of the antibody).

Cancer patients

Five patients with advanced cancer (colorectal cancer, cholangiocarcinoma, or NSCLC) with elevated serum CRP levels were dosed with either 80 mg or 160 mg of Abl. Serum CRP levels were greatly reduced in these patients (Figure 26A). The reduction in serum CRP levels was rapid, 90% reduction occurred within one week of Abl administration and lasted at least until the final measurement was made (up to twelve weeks). Figure 26B shows the CRP levels of two representative individuals. In those individuals, CRP levels decreased below the normal reference range (less than 5 - 6 mg / 1) within one week. Therefore, the administration of Abl to patients with advanced cancer can cause rapid and sustained suppression of serum CRP levels.

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IMPI

Example 21 Abl improved muscle strength, improved- weight and reduced fatigue in patients with advanced cancer

Introduction

Weight loss and fatigue (and the muscle weakness that comes with them) are very common symptoms in patients with advanced forms of cancer, and these symptoms may worsen as the cancer progresses. Fatigue, weight loss and muscle weakness can have considerable negative effects on the recovery of patients with advanced forms of cancer, for example, altering lifestyles and relationships and affecting the will or ability of patients to continue treatments against cancer. Known methods of addressing fatigue, weight loss, and muscle weakness include regular training and exercise routines, methods to conserve patient energy, and treatments that address anemia-induced fatigue and muscle weakness. However, there remains a need in the art for methods and / or treatments to improve fatigue, weight loss, and muscle weakness in cancer patients.

Methods

Four patients with advanced forms of cancer (colorectal cancer (2), NSCLC (1), cholangiocarcinoma (1)) received a single 1-hour intravenous (IV) infusion of 80 mg or 160 mg of the monoclonal antibody Abl. No doses were administered

526

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proof.

Patients were evaluated before dose administration and thereafter for at least 6 weeks after dose. At the time of each assessment, patients were screened for the following: a.) Any change in weight, b.) Fatigue measured using the Facit-F fatigue subscale questionnaire, a medically recognized test to assess fatigue (See, for example, Celia, D.,

Lai, JS, Chang, CH, Peterman, A., & Slavin, M. (2002).

Fatigue in cancer patients compared with fatigue in the general population. Cancer, 94 (2), 528-538; Celia, D., Eton, DT,

Lai, F JS., Peterman, AH & Merkel, DE (2002). Combining anchor and distribution based methods to derive minimal clinically important differences on the Functional Assessment of Cancer Therapy anemia and fatigue scales. Journal of Pain &

Symptom Management, 24 (6) 547-561.) And grip strength (a medically recognized test to assess muscle strength, typically using a grip dynamometer).

Results

Change in weight

The averaged data for both dose concentrations (80 mg and 160 mg) of the monoclonal antibody Abl demonstrated an increase of approximately 2 kilograms of weight per patient over the 6 week period (Figure 29).

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Fatigue

The averaged data for both dose concentrations (80 mg and 160 mg) of the monoclonal antibody Abl demonstrated an increase in the mean Facit-F FS subscale result of at least approximately 10 points in the patient population over the period of 6 weeks (Figure 30).

Gripping force

The averaged data for both dose concentrations (80 mg and 160 mg) of the monoclonal antibody Abl demonstrated an increase in mean grip strength of at least approximately 10 percent in the patient population over the 6-week period (Figure 31). .

Example 22 Abl increases plasma albumin concentration in patients with advanced cancer

Introduction

Serum albumin concentrations are recognized as predictive indicators of survival and / or recovery success in cancer patients. Hypoalbuminenia has a strong correlation with poor patient outcomes in various forms of cancer. For example, in one study, no patient undergoing systemic chemotherapy for metastatic pancreatic adenocarcinoma and with serum albumin levels less than 3.5 g / dL responded satisfactorily to systemic chemotherapy (Fujishiro, M., et al.,

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MEXICAN INSTITUTE

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INDUSTRIAL

Hepatogastroenterology, 47 (36): 1744-46 (2000)). The authors conclude that [patients with ... hypoalbuminemia ... may be inappropriate candidates for systemic chemotherapy and could be treated with other experimental approaches or palliative care. Id

Similarly, Sénior and Maroni express that [t] he recent appreciation that hypoalbuminemia is the most powerful indicator of mortality in end-stage renal disease highlights the critical importance of ensuring adequate protein intake for this patient population. . (JR Senior and BJ Maroni, Am. Soc. Nutr. Sci., 129: 313S-314S (1999)).

In at least one study, attempts to rectify hypoalbuminemia in 27 patients with metastatic cancer by infusing 20 g daily intravenous albumin until normal serum albumin levels were reached (> 3.5 g / dL) were not very successful . The authors mention that [l] albumin infusion in patients with advanced stage cancer has limited value in clinical practice. Patients with PS 4 and hypoalbuminemia have a worse prognosis. (Demirkazik,

A., et al., Proc. Am. Soc. Clin. Oncol., 21: Abstr 2892 (2002)).

Accordingly, there remains a need in the art for methods and / or treatments that improve serum albumin concentrations in cancer patients and that address hypoalbuminemic states in cancer patients,

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MEXICAN INSTITUTE particularly, those with advanced cancers ^ ^ ^ ^ ¿

Methods

Patients were evaluated before dose administration and thereafter for at least 6 weeks after dose. At the time of each evaluation, the plasma albumin concentration in the patients was analyzed.

Results

The averaged data for both dosage concentrations (80 mg and 160 mg) of the monoclonal antibody Abl demonstrated an increase of approximately 5 g / L in plasma albumin concentration per patient over the 6 week period (Figure 33).

Example 23 Abl increases hemoglobin in patients with advanced cancer

The Abl antibody was dosed to 93 individuals with non-small cell lung carcinoma at 80 mg, 160 mg or 320 mg Abl in phosphate buffered saline. The placebo-treated group of 31 individuals with non-small cell lung carcinoma

530

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A subgroup of the study population began the study with low hemoglobin levels, defined as a reference hemoglobin concentration below 11 g / 1. The average hemoglobin concentration increased above 11 g / 1 after eight weeks for those who received the Abl antibody in doses of 160 mg and 320 mg, while the average hemoglobin concentration of those who received Abl antibody in a dose of 80 mg or placebo did not increase above 11 g / 1 after eight weeks (Fig.

40) .

These results further demonstrate some of the beneficial effects of Abl administration to individuals with chronic diseases. Because IL-6 is the main cytokine responsible for anemia in chronic diseases (including anemia related to cancer), neutralization of IL-6 by Abl increases the concentration

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IMPI of hemoglobin in these individuals. ΙΝ3Τ,™ ΰ? Sedad

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Example 24 Abl increases hemoglobin in patients with rheumatoid arthritis

Hemoglobin levels were analyzed in patients with rheumatoid arthritis during treatment with Abl antibody. The Abl antibody was dosed at 80 individuals, 160 mg or 320 mg in phosphate buffered saline to 94 individuals with

I rheumatoid arthritis. The placebo-treated group of 33 individuals with rheumatoid arthritis was dosed only with phosphate buffered saline. Blood samples were drawn just before dosing (week zero) and at week one, two, three, four, six, eight, ten, twelve and sixteen and the hemoglobin concentration was determined. The average hemoglobin concentration increased for those receiving the Abl antibody while the average hemoglobin concentration for those receiving placebo did not significantly increase after sixteen weeks when compared to the concentration just before dosing (week zero) (Figure 41 ).

These results further demonstrate some of the beneficial effects of Abl administration to individuals with chronic diseases. Because IL-6 is the primary cytokine responsible for anemia of chronic disease (including cancer-related anemia),

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INDUSTRIAL

Neutralization of IL-6 by Abl increases the concentration of hemoglobin.

Ejeaplo 25 Abl improved weight and reduced fatigue in patients with advanced cancer

Introduction

Weight loss and fatigue are very common symptoms in patients with advanced forms of cancer, and these symptoms may worsen as the cancer progresses. Fatigue and weight loss can have considerable negative effects on the recovery of patients with advanced forms of cancer, for example by altering lifestyles and relationships and affecting patients' willingness or ability to continue cancer treatments. Known methods of addressing fatigue and weight loss include regular training and exercise routines, methods to conserve patient energy, and treatments that address anemia-induced fatigue. However, there remains a need in the art for methods and / or treatments to improve fatigue and weight loss in cancer patients.

Methods

One hundred twenty-four non-small cell lung cancer (NSCLC) patients were divided into 4 treatment groups. Patients in one group received a 1-hour intravenous (IV) infusion of placebo (n = 31), 80 mg (n = 29), 160 mg (n = 32), or 320

533

monoclonal antibody Abl mg (n = 32) every 8 weeks for weeks for a total of 3 doses.

Patients were evaluated before dose administration and thereafter for at least 12 weeks after dose. At the time of each assessment, patients were screened for the following: a.) Any change in weight, and b.) Fatigue measured using the Facit-F fatigue subscale questionnaire, a medically recognized test to assess fatigue (See, eg, Celia, D., Lai,

JS, Chang, CH, Peterman, A., & Slavin, M. (2002). Fatigue in cancer patients compared with fatigue in the general population. Cancer, 94 (2), 528-538; Celia, D., Eton, DT, Lai, F JS., Peterman, AH & Merkel, DE (2002). Combining anchor and distribution based methods to derive minimal clinically important differences on the Functional Assessment of Cancer Therapy anemia and fatigue scales. Journal of Pain &

Symptom Management, 24 (6) 547-561.).

Results

Change in weight

The averaged data of change in weight for each dose concentration group (placebo, 80 mg, 160 mg, and 320 mg) of the monoclonal antibody Abl over 12 weeks are plotted in Figure 42. The average percentage change in body weight of each dose concentration is plotted in Figure 43. The average lean body mass data for the groups of

534

Dose concentrations are plotted in Figure 44.

Fatigue

Fatigue averaged for each dose concentration group (placebo, 80 mg, 160 mg, and 320 mg) of the monoclonal antibody Abl demonstrated increases in subscale score

Average Facit-F FS for some of the dose concentration groups in the patient population over a period of 8 weeks (Figure 45). Changing the score is subscale

Reference Facit-F is plotted in Figure 46.

SEQUENCE LISTING

The biological sequences referred to herein are provided below:

SEQ ID NO: 1

VPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENNLNL

PKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQ

KKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKEFLQSSLRALRQM

SEQ ID NO: 2

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKPG

QRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNAFGGG

TEWVKRTVAAPSVFIFPPSDEQLKSGTASWCLLNN

SEQ ID NO: 3

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYVTWVRQAPGKGL

EWIGIIYGSDETAYATWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNLW

GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK

535

IMPI

OOUANO INSTITUTE OE LA Ra /; ... ·· Π'Λϋ

INDUSIRUL

SEQ ID NO: 4

QASQSINNELS

SEQ ID NO: 5

RASTLAS

SEQ ID NO: 6

QQGYSLRNIDNA

SEQ ID NO: 7

NYYVT

SEQ ID NO: 8

IIYGSDETAYATWAIG

SEQ ID NO: 9

DDSSDWDAKFNL

SEQ ID NO: 10

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAACCAGGG

CAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCTTTCGGCGGAGGG

ACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGAT

GAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTT

SEQ ID NO: 11

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

536

GAATGGATCGGAATCATTTATGGTAGTGATGAAACGGCCTACGCGACCTGGGCGATAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCCGCGGAC

ACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAACTTGTGG

GGCCAAGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTG

GCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG

SEQ ID NO: 12

CAGGCCAGTCAGAGCATTAACAATGAATTATCC

SEQ ID NO: 13

AGGGCATCCACTCTGGCATCT

SEQ ID NO: 14

CAACAGGGTTATAGTCTGAGGAATATTGATAATGCT

SEQ ID NO: 15

AACTACTACGTGACC

SEQ ID NO: 16

ATCATTTATGGTAGTGATGAAACGGCCTACGCGACCTGGGCGATAGGC

SEQ ID NO: 17

GATGATAGTAGT GACT GGGAT GCAAAATT TAACT TG

SEQ ID NO: 18

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATWA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNL

SEQ ID NO: 19

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATSA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNL

SEQ ID NO: 20

IQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFSG

537

SGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNA

SEQ ID NO: 21 --- -MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTINCQASETIYSWLSWYQQKPG

QPPKLLIYQASDLASGVPSRFSGSGAGTEYTLTISGVQCDDAATYYCQQGYSGSNVDNVFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAK

SEQ ID NO: 22

METGLRWLLLVAVLKGVQCQEQLKESGGRLVTPGTPLTLTCTASGFSLNDHAMGWVRQAPGKG

LEYIGFINSGGSARYASWAEGRFTISRTSTTVDLKMTSLTTEDTATYFCVRGGAVWSIHSFDP

WGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK

SEQ ID NO: 23

QASETIYSWLS

SEQ ID NO: 24

QASDLAS

SEQ ID NO: 25

QQGYSGSNVDNV

SEQ ID NO: 26

DHAMG

SEQ ID NO: 27

FINSGGSARYASWAEG

SEQ ID NO: 28

GGAVWSIHSFDP

SEQ ID NO: 29

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGGCCAGTGAGACCATTTACAGTTGGTTATCCTGGTATCAGCAGAAGCCAGGG

538

INSTITUTO MEXICANO DE LA, 'ROPltüAD INDUSTRIAL

CAGCCTCCCAAGCTCCTGATCTACCAGGCATCCGATCTGGCATCTGGGGTCCCATCGCGATTC

AGCGGCAGTGGGGCTGGGACAGAGTACACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTGGTAGTAATGTTGATAATGTTTTCGGCGGAGGGG

ACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGAT

GAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAG

GCCAAAG

SEQ ID NO: 30

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGGAG

CAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTTACCTGCACA

GCCTCTGGATTCTCCCTCAATGACCATGCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTGGAATACATCGGATTCATTAATAGTGGTGGTAGCGCACGCTACGCGAGCTGGGCAGAAGGC

CGATTCACCATCTCCAGAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAG

GACACGGCCACCTATTTCTGTGTCAGAGGGGGTGCTGTTTGGAGTATTCATAGTTTTGATCCC

TGGGGCCCAGGGACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCC

CTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAG

SEQ ID NO: 31

CAGGCCAGTGAGACCATTTACAGTTGGTTATCC

SEQ ID NO: 32

CAGGCATCCGATCTGGCATCT

SEQ ID NO: 33

CAACAGGGT TATAGTGGTAGTAAT GTT GATAAT GTT

SEQ ID NO: 34

GACCATGCAATGGGC

SEQ ID NO: 35

TTCATTAATAGTGGTGGTAGCGCACGCTACGCGAGCTGGGCAGAAGGC

539 £ „i

MEXICAN INSTITUTE \ Ñ'Ñ DE LA PROPIEDAD V \ - - INDUSTRIAL

SEQ ID NO: 36

GGGGGTGCTGTTTGGAGTATTCATAGTTTTGATCCC

SEQ ID NO: 37

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQASQSVYDNNYLSWFQQK

PGQPPKLLIYGASTLASGVPSRFVGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDNAFGG

GTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF

SEQ ID NO: 38

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSVYYMNWVRQAPGKGL

EWIGFITMSDNINYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARSRGWGTMGRLDLW

GPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK

SEQ ID NO: 39

QASQSVYDNNYLS

SEQ ID NO: 40

GASTLAS

SEQ ID NO: 41

AGVYDDDSDNA

SEQ ID NO: 42 vyYmn

SEQ ID NO: 43

FITMSDNINYASWAKG

SEQ ID NO: 44

SRGWGTMGRLDL

SEQ ID NO: 45

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGC

540

ATCAGTTGCCAGGCCAGTCAGAGTGTTTATGACAACAACTACTTATCCTGGTTTCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCGTGGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACAGACGTGCAGTGTGAC

GATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATAATGCCTTCGGCGGA

GGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCT

GATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCT

SEQ ID NO: 46

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACCCCTGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAGTGTCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGATTCATTACAATGAGTGATAATATAAATTACGCGAGCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTACAATGGGTCGGTTGGATCTCTGG

GGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTG

GCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG

SEQ ID NO: 47

CAGGCCAGTCAGAGTGTTTATGACAACAACTACTTATCC

SEQ ID NO: 48

GGTGCATCCACTCTGGCATCT

SEQ ID NO: 49

GCAGGCGTTTATGATGATGATAGTGATAATGCC

SEQ ID NO: 50

GTCTACTACATGAAC

SEQ ID NO: 51

TTCATTACAATGAGTGATAATATAAATTACGCGAGCTGGGCGAAAGGC

541

SEQ ID NO: 52

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MEXICO INSTITUTE · '. ·) OF THE EROHEOAO

INDUSTRIAL

AGTCGTGGCTGGGGTACAATGGGTCGGTTGGATCTC

SEQ ID NO: 53

MDTRAPTQLLGLLLLWLPGAICDPVLTQTPSPVSAPVGGTVSISCQASQSVYENNYLSWFQQK

PGQPPKLLIYGASTLDSGVPSRFKGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDDAFGG

GTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN

SEQ ID NO: 54

METGLRWLLLVAVLKGVQCQEQLKESGGGLVTPGGTLTLTCTASGFSLNAYYMNWVRQAPGKG

LEWIGFITLNNNVAYANWAKGRFTFSKTSTTVDLKMTSPTPEDTATYFCARSRGWGAMGRLDL

WGHGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK

SEQ ID NO: 55

QASQSVYENNYLS

SEQ ID NO: 56

GASTLDS

SEQ ID NO: 57

AGVYDDDSDDA

SEQ ID NO: 58

AYYMN

SEQ ID NO: 59

FITLNNNVAYANWAKG

SEQ ID NO: 60

SRGWGAMGRLDL

SEQ ID NO: 61

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCATA

TGTGACCCTGTGCTGACCCAGACTCCATCTCCCGTATCTGCACCTGTGGGAGGCACAGTCAGC

542

ATCAGTTGCCAGGCCAGTCAGAGTGTTTATGAGAACAACTATTTATCCTGGTTTCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGATTCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATTACAGACGTGCAGTGTGAC

GATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATGATGCCTTCGGCGGA

GGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCT

GATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTT

SEQ ID NO: 62

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGTCAGGAG

CAGCTGAAGGAGTCCGGAGGAGGCCTGGTAACGCCTGGAGGAACCCTGACACTCACCTGCACA

GCCTCTGGATTCTCCCTCAATGCCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTGGAATGGATCGGATTCATTACTCTGAATAATAATGTAGCTTACGCGAACTGGGCGAAAGGC

CGATTCACCTTCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACACCCGAG

GACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTGCAATGGGTCGGTTGGATCTC

TGGGGCCATGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCC

CTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG

SEQ ID NO: 63

CAGGCCAGT CAGAGT GT T TAT GAGAACAACTATTTATCC

SEQ ID NO: 64

GGTGCATCCACTCTGGATTCT

SEQ ID NO: 65

GCAGGCGTTTATGATGATGATAGTGATGATGCC

SEQ ID NO: 66

GCCTACTACATGAAC

SEQ ID NO: 67

TTCATTACTCTGAATAATAATGTAGCTTACGCGAACTGGGCGAAAGGC

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Mexican Institute of Industrial Property

SEQ ID NO: 68

AGTCGTGGCTGGGGTGCAATGGGTCGGTTGGATCTC

SEQ ID NO: 69

MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSPVSAAVGGTVTINCQASQSVDDNNWLGWYQQK

RGQPPKYLIYSASTLASGVPSRFKGSGSGTQFTLTISDLECDDAATYYCAGGFSGNIFAFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF

SEQ ID NO: 70

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGL

EWIGIIGGFGTTYYATWAKGRFTISKTSTTVDLRITSPTTEDTATYFCARGGPGNGGDIWGQG

TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD

SEQ ID NO: 71

QASQSVDDNNWLG

SEQ ID NO: 72

SASTLAS

SEQ ID NO: 73

AGGFSGNIFA

SEQ ID NO: 74

SYAMS

SEQ ID NO: 75

IIGGFGTTYYATWAKG

SEQ ID NO: 76

GGPGNGGDI

SEQ ID NO: 77

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCCAAGTGCTGACCCAGACTCCATCGCCTGTGTCTGCAGCTGTGGGAGGCACAGTCACC

544

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INSTITUTE ΜίΧΙΓΛ NOT OF THE INDUSTRIAL mCHtoñ

ATCAACTGCCAGGCCAGTCAGAGTGTTGATGATAACAACTGGTTAGGCTGGTATCAGCAGAAA

CGAGGGCAGCCTCCCAAGTACCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGAC

GATGCTGCCACTTACTACTGTGCAGGCGGTTTTAGTGGTAATATCTTTGCTTTCGGCGGAGGG

ACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGAT

GAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCT

SEQ ID NO: 78

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGCTTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGAAAGGGGCTG

GAGTGGATCGGAATCATTGGTGGTTTTGGTACCACATACTACGCGACCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAGAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGGTGGTCCTGGTAATGGTGGTGACATCTGGGGCCAAGGG

ACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCC

TCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACT

SEQ ID NO: 79

CAGGCCAGT CAGAGT GTT GAT GATAACAACTGGT TAGGC

SEQ ID NO: 80

TCTGCATCCACTCTGGCATCT

SEQ ID NO: 81

GCAGGCGGTTTTAGTGGTAATATCTTTGCT

SEQ ID NO: 82

AGCTATGCAATGAGC

SEQ ID NO: 83

ATCATTGGTGGTTTTGGTACCACATACTACGCGACCTGGGCGAAAGGC

545 ίΡΙ

INSTITUTE ME. ' ICA'Oj de la Piro ielv u INDUSTRIAL

SEQ ID NO: 84

GGTGGTCCTGGTAATGGTGGTGACATC '

SEQ ID NO: 85

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSVPVGGTVTIKCQSSQSVYNNFLSWYQQKP

GQPPKLLIYQASKLASGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNAFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF

SEQ ID NO: 86

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGIDLSDYAMSWVRQAPGKGL

EWIGIIYAGSGSTWYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARDGYDDYGDFDRL

DLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD

SEQ ID NO: 87

QSSQSVYNNFLS

SEQ ID NO: 88

QASKLAS

SEQ ID NO: 89

LGGYDDDADNA

SEQ ID NO: 90

DYAMS

SEQ ID NO: 91

IIYAGSGSTWYASWAKG

SEQ ID NO: 92

DGYDDYGDFDRLDL

SEQ ID NO: 93

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGTACCTGTGGGAGGCACAGTCACC

546

INSTITUTO ÁtEXÍC /; ¡í>

D £ THE PAPY »

INDUSTRIAL

ATCAAGTGCCAGTCCAGTCAGAGTGTTTATAATAATTTCTTATCGTGGTATCAGCAGAAACCA

GGGCAGCCTCCCAAGCTCCTGATCTACCAGGCATCCAAACTGGCATCTGGGGTCCCAGATAGG

TTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGAT

GCTGCCACTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCTTTCGGCGGAGGGG

ACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGAT

GAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTC

SEQ ID NO: 94

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACGCTCACCTGCACAGTC

TCTGGAATCGACCTCAGTGACTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATCATTTATGCTGGTAGTGGTAGCACATGGTACGCGAGCTGGGCGAAAGGC

CGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAG

GACACGGCCACCTATTTCTGTGCCAGAGATGGATACGATGACTATGGTGATTTCGATCGATTG

GATCTCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTC

TTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTC

AAGGACT

SEQ ID NO: 95

CAGTCCAGTCAGAGTGTTTATAATAATTTCTTATCG

SEQ ID NO: 96

CAGGCATCCAAACTGGCATCT

SEQ ID NO: 97

CTAGGCGGTTATGATGATGATGCTGATAATGCT

SEQ ID NO: 98

GACTATGCAATGAGC

SEQ ID NO: 99

547

ATCATTTATGCTGGTAGTGGTAGCACATGGTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 100 -

GATGGATACGATGACTATGGTGATTTCGATCGATTGGATCTC

SEQ ID NO: 101

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKSG

QRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNAFGGG

TEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF

SEQ ID NO: 102

METGLRWLLLVAVLSGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYMTWVRQAPGKGL

EWIGMIYGSDETAYANWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNLW

GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK

SEQ ID NO: 103

QASQSINNELS

SEQ ID NO: 104

RASTLAS

SEQ ID NO: 105

QQGYSLRNIDNA

SEQ ID NO: 106

NYYMT

SEQ ID NO: 107

MIYGSDETAYANWAIG

SEQ ID NO: 108

DDSSDWDAKFNL

SEQ ID NO: 109

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

548

IMP

IND'JS I EUL - -

TGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAAATGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAATCAGGG

CAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCTTTCGGCGGAGGG

ACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGAT

GAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTC

SEQ ID NO: 110

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCTCAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAGTAACTACTACATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATGATTTATGGTAGTGATGAAACAGCCTACGCGAACTGGGCGATAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCCGCGGAC

ACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAACTTGTGG

GGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTG

GCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG

SEQ ID NO: 111

CAGGCCAGT CAGAGCAT TAACAAT GAATTATCC

SEQ ID NO: 112

AGGGCATCCACTCTGGCATCT

SEQ ID NO: 113

CAACAGGGT TATAGT CT GAGGAATATTGATAATGCT

SEQ ID NO: 114

AACTACTACATGACC

SEQ ID NO: 115

549

ΙΜ

INSTITUTE ME.'UCANO OF INDUSTRIAL PROPERTY

ATGATTTATGGTAGTGATGAAACAGCCTACGCGAACTGGGCGATAGGC u lAL

SEQ ID NO: 116 ~ —--- GATGATAGTAGTGACTGGGATGCAAAATTTAACTTG

SEQ ID NO: 117

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYMTWVRQAPGKGLEWVGMIYGSDETAYANWA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSS DWDAKFNL

SEQ ID NO: 118

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYMTWVRQAPGKGLEWVGMIYGSDETAYANSA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNL

SEQ ID NO: 119

DIQMTQSPSTLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFS

GSGSGTEFTLTISSLQPDDFATYYCQQGYSLRNIDNA

SEQ ID NO: 120

IIYGSDETAYATSAIG

SEQ ID NO: 121

MIYGSDETAYANSAIG

SEQ ID NO: 122

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQSSQSVGNNQDLSWFQQR

PGQPPKLLIYEISKLESGVPSRFSGSGSGTHFTLTISGVQCDDAATYYCLGGYDDDADNA

SEQ ID NO: 123

METGLRWLLLVAVLKGVQCHSVEESGGRLVTPGTPLTLTCTVSGFSLSSRTMSWVRQAPGKGL

EWIGYIWSGGSTYYATWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARLGDTGGHAYATRL

NL

SEQ ID NO: 124

QSSQSVGNNQDLS

550

SEQ ID NO: 125

EISKLES ~

SEQ ID NO: 126

LGGYDDDADNA

SEQ ID NO: 127

SRTMS

SEQ ID NO: 128

YIWSGGSTYYATWAKG

SEQ ID NO: 129

LGDTGGHAYATRLNL

SEQ ID NO: 130

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCAGCCGTGCTGACCCAGACACCATCACCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAGTTGCCAGTCCAGTCAGAGTGTTGGTAATAACCAGGACTTATCCTGGTTTCAGCAGAGA

CCAGGGCAGCCTCCCAAGCTCCTGATCTACGAAATATCCAAACTGGAATCTGGGGTCCCATCG

CGGTTCAGCGGCAGTGGATCTGGGACACACTTCACTCTCACCATCAGCGGCGTACAGTGTGAC

GATGCTGCCACTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCT

SEQ ID NO: 131

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCACTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGATTCTCCCTCAGTAGTCGTACAATGTCCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAGTGGATCGGATACATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGATTGGGCGATACTGGTGGTCACGCTTATGCTACTCGCTTA

AATCTC

551

IΜ> τ

INjTM oto MEXICANO

SEQ ID NO: 132 industrial

CAGTCCAGTCAGAGTGTTGGTAATAACCAGGACTTATCC ———

SEQ ID NO: 133

GAAATATCCAAACTGGAATCT

SEQ ID NO: 134

CTAGGCGGTTATGATGATGATGCTGATAATGCT

SEQ ID NO: 135

AGTCGTACAATGTCC

SEQ ID NO: 136

TACATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAAGGC

SEQ ID NO: 137

TTGGGCGATACTGGTGGTCACGCTTATGCTACTCGCTTAAATCTC

SEQ ID NO: 138

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSSVSAAVGGTVSISCQSSQSVYSNKYLAWYQQK

PGQPPKLLIYWTSKLASGAPSRFSGSGSGTQFTLTISGVQCDDAATYYCLGAYDDDADNA

SEQ ID NO: 139

METGLRWLLLVAVLKGVQCQSVEESGGRLVKPDETLTLTCTASGFSLEGGYMTWVRQAPGKGL

EWIGISYDSGSTYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCVRSLKYPTVTSDDL

SEQ ID NO: 140

QSSQSVYSNKYLA

SEQ ID NO: 141

WTSKLAS

SEQ ID NO: 142

LGAYDDDADNA

SEQ ID NO: 143

552

GGYMT

SEQ ID NO: 144 ~

ISYDSGSTYYASWAKG

SEQ ID NO: 145

SLKYPTVTSDDL

SEQ ID NO: 146

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCAGCCGTGCTGACCCAGACACCATCGTCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGC

ATCAGTTGCCAGTCCAGTCAGAGTGTTTATAGTAATAAGTACCTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTACTGGACATCCAAACTGGCATCTGGGGCCCCATCA

CGGTTCAGCGGCAGTGGATCTGGGACACAATTCACTCTCACCATCAGCGGCGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTCTAGGCGCTTATGATGATGATGCTGATAATGCT

SEQ ID NO: 147

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAAGAGTCCGGGGGTCGCCTGGTCAAGCCTGACGAAACCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTGGAGGGCGGCTACATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATCAGTTATGATAGTGGTAGCACATACTACGCGAGCTGGGCGAAAGGCCGA

TTCACCATCTCCAAGACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAG

GACACGGCCACCTATTTCTGCGTCAGATCACTAAAATATCCTACTGTTACTTCTGATGACTTG

SEQ ID NO: 148

CAGTCCAGTCAGAGTGTTTATAGTAATAAGTACCTAGCC

SEQ ID NO: 149

TGGACATCCAAACTGGCATCT

SEQ ID NO: 150

CTAGGCGCTTATGATGATGATGCTGATAATGCT

553

INSTITUTE .MtXICAW, Say LA PP.OPIÍD. '. Ü

INOUSTMIAL

SEQ ID NO: 151

GGCGGCTACATGACC

SEQ ID NO: 152

ATCAGTTATGATAGTGGTAGCACATACTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 153

TCACTAAAATATCCTACTGTTACTTCTGATGACTTG

SEQ ID NO: 154

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQSSQSVYNNNDLAWYQQK

PGQPPKLLIYYASTLASGVPSRFKGSGSGTQFTLTISGVQCDDAAAYYCLGGYDDDADNA

SEQ ID NO: 155

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGLSLSSNTINWVRQAPGKGL

EWIGYIWSGGSTYYASWVNGRFTISKTSTTVDLKITSPTTEDTATYFCARGGYASGGYPYATR

LDL

SEQ ID NO: 156

QSSQSVYNNNDLA

SEQ ID NO: 157

YASTLAS

SEQ ID NO: 158

LGGYDDDADNA

SEQ ID NO: 159

SNTIN

SEQ ID NO: 160

YIWSGGSTYYASWVNG

SEQ ID NO: 161

GGYASGGYPYATRLDL

554

IMPI

SEQ ID NO: 162 ________—

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCAGCCGTGCTGACCCAGACACCATCACCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAGTTGCCAGTCCAGTCAGAGTGTTTATAATAATAACGACTTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCTAAACTCCTGATCTATTATGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGAC

GATGCTGCCGCTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCT

SEQ ID NO: 163

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTA

TCTGGATTATCCCTCAGTAGCAATACAATAAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAGTGGATCGGATACATTTGGAGTGGTGGTAGTACATACTACGCGAGCTGGGTGAATGGTCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGGGGGTTACGCTAGTGGTGGTTATCCTTATGCCACTCGG

TTGGATCTC

SEQ ID NO: 164

CAGTCCAGTCAGAGTGTTTATAATAATAACGACTTAGCC

SEQ ID NO: 165

TATGCATCCACTCTGGCATCT

SEQ ID NO: 166

CTAGGCGGT TAT GAT GAT GAT GCTGATAATGCT

SEQ ID NO: 167

AGCAATACAATAAAC

SEQ ID NO: 168

TACATTTGGAGTGGTGGTAGTACATACTACGCGAGCTGGGTGAATGGT

555

MCAÍ-CUYO INSTITUTE Ofc LA PROJnCYV '

IMvUSTíüÁÍ

SEQ ID NO: 169 -GGGGGTTACGCTAGTGGTGGTTATCCTTATGCCACTCGGTTGGATCTC

SEQ ID NO: 170

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSSVSAAVGGTVTINCQSSQSVYNNDYLSWYQQR

PGQRPKLLIYGASKLASGVPSRFKGSGSGKQFTLTISGVQCDDAATYYCLGDYDDDADNT

SEQ ID NO: 171

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFTLSTNYYLSWVRQAPGKG

LEWIGIIYPSGNTYCAKWAKGRFTISKTSSTTVDLKMTSPTTEDTATYFCARNYGGDESL

SEQ ID NO: 172

QSSQSVYNNDYLS

SEQ ID NO: 173

GASKLAS

SEQ ID NO: 174

LGDYDDDADNT

SEQ ID NO: 175

TNYYLS

SEQ ID NO: 176

IIYPSGNTYCAKWAKG

SEQ ID NO: 177

NYGGDESL

SEQ ID NO: 178

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCAGCCGTGCTGACCCAGACACCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGTCCAGTCAGAGTGTTTATAATAACGACTACTTATCCTGGTATCAACAGAGG

CCAGGGCAACGTCCCAAGCTCCTAATCTATGGTGCTTCCAAACTGGCATCTGGGGTCCCGTCA

556

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

CGGTTCAAAGGCAGTGGATCTGGGAAACAGTTTACTCTCACCATCAGCGGCGTGCAGTG1

GATGCTGCCACTTACTACTGTCTGGGCGATTATGATGATGATGCTGATAATACT

SEQ ID NO: 179

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACTTGCACAGTC

TCTGGATTCACCCTCAGTACCAACTACTACCTGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTAGAATGGATCGGAATCATTTATCCTAGTGGTAACACATATTGCGCGAAGTGGGCGAAAGGC

CGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACC

GAGGACACAGCCACGTATTTCTGTGCCAGAAATTATGGTGGTGATGAAAGTTTG

SEQ ID NO: 180

CAGTCCAGTCAGAGTGTTTATAATAACGACTACTTATCC

SEQ ID NO: 181

GGTGCTTCCAAACTGGCATCT

SEQ ID NO: 182

CTGGGCGATTATGATGATGATGCTGATAATACT

SEQ ID NO: 183

ACCAACTACTACCTGAGC

SEQ ID NO: 184

ATCATTTATCCTAGTGGTAACACATATTGCGCGAAGTGGGCGAAAGGC

SEQ ID NO: 185

AATTATGGTGGTGATGAAAGTTTG

SEQ ID NO: 186

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASETIGNALAWYQQKSG

QPPKLLIYKASKLASGVPSRFKGSGSGTEYTLTISDLECADAATYYCQWCYFGDSV

SEQ ID NO: 187

557

METGLRWLLLVTVLKGVQCQEQLVESGGGLVQPEGSLTLTCTASGFDFSSGYYMCWVRQAPGK

GLEWIACIFTITTNTYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYLCARGIYSDNNYY

TO THE

SEQ ID NO: 188

QASETIGNALA

SEQ ID NO: 189

KASKLAS

SEQ ID NO: 190

QWCYFGDSV

SEQ ID NO: 191

SGYYMC

SEQ ID NO: 192

CIFTITTNTYYASWAKG

SEQ ID NO: 193

GIYSDNNYYAL

SEQ ID NO: 194

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTGAGACCATTGGCAATGCATTAGCCTGGTATCAGCAGAAATCAGGG

CAGCCTCCCAAGCTCCTGATCTACAAGGCATCCAAACTGGCATCTGGGGTCCCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAATGGTGTTATTTTGGTGATAGTGTT

SEQ ID NO: 195

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCACTGTGCTCAAAGGTGTCCAGTGTCAGGAG

CAGCTGGTGGAGTCCGGGGGAGGCCTGGTCCAGCCTGAGGGATCCCTGACACTCACCTGCACA

558

IMPI

GCCTCTGGATTCGACTTCAGTAGCGGCTACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAGTGGATCGCGTGTATTTTCACTATTACTACTAACACTTACTACGCGAGCTGGGCG

AAAGGCCGATTCACCATCTCCAAGACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTG

ACAGCCGCGGACACGGCCACCTATCTCTGTGCGAGAGGGATTTATTCTGATAATAATTATTAT

GCCTTG

SEQ ID NO: 196

CAGGCCAGTGAGACCATTGGCAATGCATTAGCC

SEQ ID NO: 197

AAGGCATCCAAACTGGCATCT

SEQ ID NO: 198

CAATGGTGTTATTTTGGTGATAGTGTT

SEQ ID NO: 199

AGCGGCTACTACATGTGC

SEQ ID NO: 200

TGTATTTTCACTATTACTACTAACACTTACTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 201

GGGATTTATTCTGATAATAATTATTATGCCTTG

SEQ ID NO: 202

MDTRAPTQLLGLLLLWLPGARCDWMTQTPASVEAAVGGTVTIKCQASESIGNALAWYQQKPG

QPPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISGVQCADAAAYYCQWCYFGDSV

SEQ ID NO: 203

METGLRWLLLVAVLKGVQCQQQLVESGGGLVKPGASLTLTCKASGFSFSSGYYMCWVRQAPGK

GLESIACIFTITDNTYYANWAKGRFTISKPSSPTVTLQMTSLTAADTATYFCARGIYSTDNYY

TO THE

SEQ ID NO: 204

559

QASESIGNALA

IMPI

INSTITUTO MrXl-IANO Ds' -A PSOI'IsPAD INCUSTiliAL

SEQ ID NO: 205 ---— -

KASTLAS

SEQ ID NO: 206

QWCYFGDSV

SEQ ID NO: 207

SGYYMC

SEQ ID NO: 208

CIFTITDNTYYANWAKG

SEQ ID NO: 209

GIYSTDNYYAL

SEQ ID NO: 210

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTGAGAGCATTGGCAATGCATTAGCCTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTC

AGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGCCGATGCT

GCCGCTTACTACTGTCAATGGTGTTATTTTGGTGATAGTGTT

SEQ ID NO: 211

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGCAG

CAGCTGGTGGAGTCCGGGGGAGGCCTGGTCAAGCCGGGGGCATCCCTGACACTCACCTGCAAA

GCCTCTGGATTCTCCTTCAGTAGCGGCTACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAGTCGATCGCATGCATTTTTACTATTACTGATAACACTTACTACGCGAACTGGGCG

AAAGGCCGATTCACCATCTCCAAGCCCTCGTCGCCCACGGTGACTCTGCAAATGACCAGTCTG

ACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGGGGGATTTATTCTACTGATAATTATTAT

560

MEXJC INSTITUTE .-. OU OF PROPERTY

INDUSTRIAL

GCCTTG

SEQ ID NO: 212

CAGGCCAGTGAGAGCATTGGCAATGCATTAGCC

SEQ ID NO: 213

AAGGCATCCACTCTGGCATCT

SEQ ID NO: 214

CAATGGTGTTATTTTGGTGATAGTGTT

SEQ ID NO: 215

AGCGGCTACTACATGTGC

SEQ ID NO: 216

TGCATTTTTACTATTACTGATAACACTTACTACGCGAACTGGGCGAAAGGC

SEQ ID NO: 217

GGGATTTATTCTACTGATAATTATTATGCCTTG

SEQ ID NO: 218

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASQSVSSYLNWYQQKPG

QPPKLLIYRASTLESGVPSRFKGSGSGTEFTLTISDLECADAATYYCQCTYGTSSSYGAA

SEQ ID NO: 219

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGISLSSNAISWVRQAPGKGL

EWIGIISYSGTTYYASWAKGRFTISKTSSTTVDLKITSPTTEDTATYFCARDDPTTVMVMLIP

FGAGMDL

SEQ ID NO: 220

QASQSVSSYLN

SEQ ID NO: 221

RASTLES

SEQ ID NO: 222

561

I

ί ν. ·? ¿

Iva k

QCTYGTSSSYGAA

SEQ ID NO: 223

SNAIS

SEQ ID NO: 224

IISYSGTTYYASWAKG

SEQ ID NO: 225

DDPTTVMVMLIPFGAGMDL

SEQ ID NO: 226

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTCAGAGCGTTAGTAGCTACTTAAACTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGCTCCTGATCTACAGGGCATCCACTCTGGAATCTGGGGTCCCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAATGTACTTATGGTACTAGTAGTAGTTATGGTGCTGCT

SEQ ID NO: 227

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACCGTC

TCTGGTATCTCCCTCAGTAGCAATGCAATAAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATCATTAGTTATAGTGGTACCACATACTACGCGAGCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATCACTAGTCCGACAACCGAG

GACACGGCCACCTACTTCTGTGCCAGAGATGACCCTACGACAGTTATGGTTATGTTGATACCT

TTTGGAGCCGGCATGGACCTC

SEQ ID NO: 228

CAGGCCAGTCAGAGCGTTAGTAGCTACTTAAAC

SEQ ID NO: 229

562

AGGGCATCCACTCTGGAATCT

SEQ ID NO: 230

CAATGTACTTATGGTACTAGTAGTAGTTATGGTGCTGCT

SEQ ID NO: 231

AGCAAT GCAATAAGC

SEQ ID NO: 232

ATCATTAGTTATAGTGGTACCACATACTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 233

GATGACCCTACGACAGTTATGGTTATGTTGATACCTTTTGGAGCCGGCATGGACCTC

SEQ ID NO: 234

MDTRAPTQLLGLLLLWLPGATFAQVLTQTASPVSAAVGGTVTINCQASQSVYKNNYLSWYQQK

PGQPPKGLIYSASTLDSGVPLRFSGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSSGDCYA

SEQ ID NO: 235

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPEGSLTLTCTASGFSFSSYWMCWVRQAPGKGL

EWIACIVTGNGNTYYANWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCAKAYDL

SEQ ID NO: 236

QASQSVYKNNYLS

SEQ ID NO: 237

SASTLDS

SEQ ID NO: 238

LGSYDCSSGDCYA

SEQ ID NO: 239

SYWMC

SEQ ID NO: 240

CIVTGNGNTYYANWAKG

563

SEQ ID NO: 241

AYDL

SEQ ID NO: 242

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCCAAGTGCTGACCCAGACTGCATCGCCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAACTGCCAGGCCAGTCAGAGTGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAAGGCCTGATCTATTCTGCATCGACTCTAGATTCTGGGGTCCCATTG

CGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTATGCT

SEQ ID NO: 243

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

TTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGAGGGATCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCTTCAGTAGCTACTGGATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAGTGGATCGCATGCATTGTTACTGGTAATGGTAACACTTACTACGCGAACTGGGCGAAAGGC

CGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTGACAGCC

GCGGACACGGCCACCTATTTTTGTGCGAAAGCCTATGACTTG

SEQ ID NO: 244

CAGGCCAGTCAGAGTGTTTATAAGAACAACTACTTATCC

SEQ ID NO: 245

TCTGCATCGACTCTAGATTCT

SEQ ID NO: 246

CTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTATGCT

SEQ ID NO: 247

AGCTACTGGATGTGC

SEQ ID NO: 248

564

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

TGCATTGTTACTGGTAATGGTAACACTTACTACGCGAACTGGGCGAAAGGC

SEQ ID NO: 249

GCCTATGACTTG

SEQ ID NO: 250

MDTRAPTQLLGLLLLWLPGSTFAAVLTQTPSPVSAAVGGTVSISCQASQSVYDNNYLSWYQQK

PGQPPKLLIYGASTLASGVPSRFKGTGSGTQFTLTITDVQCDDAATYYCAGVFNDDSDDA

SEQ ID NO: 251

METGLRWLLLVAVPKGVQCQSLEESGGRLVTPGTPLTLTCTLSGFSLSAYYMSWVRQAPGKGL

EWIGFITLSDHISYARWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARSRGWGAMGRLDL

SEQ ID NO: 252

QASQSVYDNNYLS

SEQ ID NO: 253

GASTLAS

SEQ ID NO: 254

AGVFNDDSDDA

SEQ ID NO: 255

AYYMS

SEQ ID NO: 256

FITLSDHISYARWAKG

SEQ ID NO: 257

SRGWGAMGRLDL

SEQ ID NO: 258

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTTCCACA

TTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGC

ATCAGTTGCCAGGCCAGTCAGAGTGTTTATGACAACAACTATTTATCCTGGTATCAGCAGAAA

565

ΙΝύ 11 i υ i ί 'fu,?. ·. * ,,. ·· J D2 LA FROPÍc:'? / · 3 INDUSTRIAL

CCAGGACAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCACGGGATCTGGGACACAGTTCACTCTCACCATCACAGACGTGCAGTGTGAC

GATGCTGCCACTTACTATTGTGCAGGCGTTTTTAATGATGATAGTGATGATGCC

SEQ ID NO: 259

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCCCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACACTC

TCTGGATTCTCCCTCAGTGCATACTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGATTCATTACTCTGAGTGATCATATATCTTACGCGAGGTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTGCAATGGGTCGGTTGGATCTC

SEQ ID NO: 260

CAGGCCAGT CAGAGTGT T TAT GACAACAACTAT T TAT CC

SEQ ID NO: 261

GGTGCATCCACTCTGGCATCT

SEQ ID NO: 262

GCAGGCGTTTTTAATGATGATAGTGATGATGCC

SEQ ID NO: 263

GCATACTATATGAGC

SEQ ID NO: 264

TTCATTACTCTGAGTGATCATATATCTTACGCGAGGTGGGCGAAAGGC

SEQ ID NO: 265

AGTCGTGGCTGGGGTGCAATGGGTCGGTTGGATCTC

SEQ ID NO: 266

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQASQSVYNNKNLAWYQQK

SGQPPKLLIYWASTLASGVSSRFSGSGSGTQFTLTVSGVQCDDAATYYCLGVFDDDADNA

566

SEQ ID NO: 267

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTASGFSLSSYSMTWVRQAPGKGL

EYIGVIGTSGSTYYATWAKGRFTISRTSTTVALKITSPTTEDTATYFCVRSLSSITFL

SEQ ID NO: 268

QASQSVYNNKNLA

SEQ ID NO: 269

WASTLAS

SEQ ID NO: 270

LGVFDDDADNA

SEQ ID NO: 271

SYSMT

SEQ ID NO: 272

VIGTSGSTYYATWAKG

SEQ ID NO: 273

SLSSITFL

SEQ ID NO: 274

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTCGCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGCGGCTGTGGGAGGCACAGTCACC

ATCAGTTGCCAGGCCAGTCAGAGTGTTTATAACAACAAAAATTTAGCCTGGTATCAGCAGAAA

TCAGGGCAGCCTCCCAAGCTCCTGATCTACTGGGCATCCACTCTGGCATCTGGGGTCTCATCG

CGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCGTCAGCGGCGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTCTAGGCGTTTTTGATGATGATGCTGATAATGCT

SEQ ID NO: 275

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAATGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCC

567

MEXICAN INSTITUTE

OF PROPERTY V

INDUSTRIAL ->

TCTGGATTCTCCCTCAGTAGCTACTCCATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATATATCGGAGTCATTGGTACTAGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGÁ '

TTCACCATCTCCAGAACCTCGACCACGGTGGCTCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGTCAGGAGTCTTTCTTCTATTACTTTCTTG

SEQ ID NO: 276

CAGGCCAGTCAGAGTGTTTATAACAACAAAAATTTAGCC

SEQ ID NO: 277

TGGGCATCCACTCTGGCATCT

SEQ ID NO: 278

CTAGGCGTTTTTGATGATGATGCTGATAATGCT

SEQ ID NO: 279

AGCTACTCCATGACC

SEQ ID NO: 280

GTCATTGGTACTAGTGGTAGCACATACTACGCGACCTGGGCGAAAGGC

SEQ ID NO: 281

AGTCTTTCTTCTATTACTTTCTTG

SEQ ID NO: 282

MDTRAPTQLLGLLLLWLPGARCAFELTQTPASVEAAVGGTVTINCQASQNIYRYLAWYQQKPG

QPPKFLIYLASTLASGVPSRFKGSGSGTEFTLTISDLECADAATYYCQSYYSSNSVA

SEQ ID NO: 283

METGLRWLLLVAVLKGVQCQEQLVESGGDLVQPEGSLTLTCTASELDFSSGYWICWVRQVPGK

GLEWIGCIYTGSSGSTFYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARGYSGFGYF

KL

SEQ ID NO: 284

QASQNIYRYLA

568

Turcinm <Τ '.) 7 .-

INSTITUTO XTXK '/ T'O DE LA PuOI-IpAO - INDUSTRIAL

SEQ ID NO: 285

LASTLAS ™ ——

SEQ ID NO: 286

QSYYSSNSVA

SEQ ID NO: 287

SGYWIC

SEQ ID NO: 288

CIYTGSSGSTFYASWAKG

SEQ ID NO: 289

GYSGFGYFKL

SEQ ID NO: 290

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCATTCGAATTGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGGCCAGTCAGAACATTTATAGATACTTAGCCTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGTTCCTGATCTATCTGGCATCTACTCTGGCATCTGGGGTCCCATCGCGGTTT

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAAAGTTATTATAGTAGTAATAGTGTCGCT

SEQ ID NO: 291

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGGAG

CAGCTGGTGGAGTCCGGGGGAGACCTGGTCCAGCCTGAGGGATCCCTGACACTCACCTGCACA

GCTTCTGAGTTAGACTTCAGTAGCGGCTACTGGATATGCTGGGTCCGCCAGGTTCCAGGGAAG

GGGCTGGAGTGGATCGGATGCATTTATACTGGTAGTAGTGGTAGCACTTTTTACGCGAGTTGG

GCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGT

CTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGAGGTTATAGTGGCTTTGGTTACTTT

AAGTTG

569

SEQ ID NO: 292

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

CAGGCCAGTCAGAACATTTATAGATACTTAGCC

SEQ ID NO: 293

CTGGCATCTACTCTGGCATCT

SEQ ID NO: 294

CAAAGT TATTATAGTAGTAATAGT GT C GCT

SEQ ID NO: 295

AGCGGCTACTGGATATGC

SEQ ID NO: 296

TGCATTTATACTGGTAGTAGTGGTAGCACTTTTTACGCGAGTTGGGCGAAAGGC

SEQ ID NO: 297

GGTTATAGTGGCTTTGGTTACTTTAAGTTG

SEQ ID NO: 298

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQASEDIYRLLAWYQQKPG

QPPKLLIYDSSDLASGVPSRFKGSGSGTEFTLAISGVQCDDAATYYCQQAWSYSDIDNA

SEQ ID NO: 299

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTASGFSLSSYYMSWVRQAPGKGL

EWIGIITTSGNTFYASWAKGRLTISRTSTTVDLKITSPTTEDTATYFCARTSDIFYYRNL

SEQ ID NO: 300

QASEDIYRLLA

SEQ ID NO: 301

DSSDLAS

SEQ ID NO: 302

QQAWSYSDIDNA

SEQ ID NO: 303

570

IMPI ·;

IhJCTITÍ ΙΤ. ^ * <- V λ μ ·>

MEXICAN INSTITUTE OF THE PRO? I £ UAl> INDUSTRIAL

SYYMS _

SEQ ID NO: 304

11TT SGNT FYASWAKG

SEQ ID NO: 305

TSDIFYYRNL

SEQ ID NO: 306

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTGAGGACATTTATAGGTTATTGGCCTGGTATCAACAGAAACCAGGG

CAGCCTCCCAAGCTCCTGATCTATGATTCATCCGATCTGGCATCTGGGGTCCCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCGCCATCAGCGGTGTGCAGTGTGACGATGCT

GCCACTTACTACTGTCAACAGGCTTGGAGTTATAGTGATATTGATAATGCT

SEQ ID NO: 307

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCGGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAGTAGCTACTACATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATCATTACTACTAGTGGTAATACATTTTACGCGAGCTGGGCGAAAGGCCGG

CTCACCATCTCCAGAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAACTTCTGATATTTTTTATTATCGTAACTTG

SEQ ID NO: 308

CAGGCCAGTGAGGACATTTATAGGTTATTGGCC

SEQ ID NO: 309

GATTCATCCGATCTGGCATCT

SEQ ID NO: 310

CAACAGGCT TGGAGT TATAGTGATAT T GATAATGCT

571

SEQ ID NO: 311

MEXICAN INSTITUTE OF THE FROPU.OAD

INDUSTRIAL

AGCTACTACATGAGC

SEQ ID NO: 312

ATCATTACTACTAGTGGTAATACATTTTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 313

ACTTCTGATATTTTTTATTATCGTAACTTG

SEQ ID NO: 314

MDTRAPTQLLGLLLLWLPGATFAAVLTQTASPVSAAVGATVTINCQSSQSVYNDMDLAWFQQK

PGQPPKLLIYSASTLASGVPSRFSGSGSGTEFTLTISGVQCDDAATYYCLGAFDDDADNT

SEQ ID NO: 315

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLTRHAITWVRQAPGKGL

EWIGCIWSGGSTYYATWAKGRFTISKTSTTVDLRITSPTTEDTATYFCARVIGDTAGYAYFTG

LDL

SEQ ID NO: 316

QSSQSVYNDMDLA

SEQ ID NO: 317

SASTLAS

SEQ ID NO: 318

LGAFDDDADNT

SEQ ID NO: 319

RHAIT

SEQ ID NO: 320

CIWSGGSTYYATWAKG

SEQ ID NO: 321

VIGDTAGYAYFTGLDL

572

IM F!

INSI'mUTD Mr / F

SEQ ID NO: 322

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGgTCGCftGGTGCCAeq ·

TTTGCAGCCGTGCTGACCCAGACTGCATCACCCGTGTCTGCCGCTGTGGGAGCCACAGTCACC

ATCAACTGCCAGTCCAGTCAGAGTGTTTATAATGACATGGACTTAGCCTGGTTTCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTCTAGGCGCTTTTGATGATGATGCTGATAATACT

SEQ ID NO: 323

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGATTCTCCCTCACTAGGCATGCAATAACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGATGCATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTCAGAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTACTTCTGTGCCAGAGTCATTGGCGATACTGCTGGTTATGCTTATTTTACGGGG

CTTGACTTG

SEQ ID NO: 324

CAGTCCAGTCAGAGTGTTTATAATGACATGGACTTAGCC

SEQ ID NO: 325

TCTGCATCCACTCTGGCATCT

SEQ ID NO: 326

CTAGGCGCTTTTGATGATGATGCTGATAATACT

SEQ ID NO: 327

AGGCATGCAATAACC

SEQ ID NO: 328

TGCATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAAGGC

573

SEQ ID NO: 329

I Μ. '

INDUS1 ¡.AL

GTCATTGGCGATACTGCTGGTTATGCTTATTTTACGGGGCTTGACTTG

SEQ ID NO: 330

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQASQSVYNWLSWYQQKPG

QPPKLLIYTASSLASGVPSRFSGSGSGTEFTLTISGVECADAATYYCQQGYTSDVDNV

SEQ ID NO: 331

METGLRWLLLVAVLKGVQCQSLEEAGGRLVTPGTPLTLTCTVSGIDLSSYAMGWVRQAPGKGL

EYIGIISSSGSTYYATWAKGRFTISQASSTTVDLKITSPTTEDSATYFCARGGAGSGGVWLLD

GFDP

SEQ ID NO: 332

QASQSVYNWLS

SEQ ID NO: 333

TASSLAS

SEQ ID NO: 334

QQGYTSDVDNV

SEQ ID NO: 335

SYAMG

SEQ ID NO: 336

IISSSGSTYYATWAKG

SEQ ID NO: 337

GGAGSGGVWLLDGFDP

SEQ ID NO: 338

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTCAGAGTGTTTATAATTGGTTATCCTGGTATCAGCAGAAACCAGGG

574

CAGCCTCCCAAGCTCCTGATCTATACTGCATCCAGTCTGGCATCTGGGGTCCCATCGCGGTTC

AGTGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAACAGGGTTATACTAGTGATGTTGATAATGTT

SEQ ID NO: 339

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGGCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGAATCGACCTCAGTAGCTATGCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATACATCGGAATCATTAGTAGTAGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGA

TTCACCATCTCACAAGCCTCGTCGACCACGGTGGATCTGAAAATTACCAGTCCGACAACCGAG

GACTCGGCCACATATTTCTGTGCCAGAGGGGGTGCTGGTAGTGGTGGTGTTTGGCTGCTTGAT

GGTTTTGATCCC

SEQ ID NO: 340

CAGGCCAGTCAGAGTGTTTATAATTGGTTATCC

SEQ ID NO: 341

ACTGCATCCAGTCTGGCATCT

SEQ ID NO: 342

CAACAGGGTTATACTAGTGATGTTGATAATGTT

SEQ ID NO: 343

AGCTATGCAATGGGC

SEQ ID NO: 344

ATCATTAGTAGTAGTGGTAGCACATACTACGCGACCTGGGCGAAAGGC

SEQ ID NO: 345

GGGGGTGCTGGTAGTGGTGGTGTTTGGCTGCTTGATGGTTTTGATCCC

SEQ ID NO: 346

MDTRAPTQLLGLLLLWLPGAKCADWMTQTPASVSAAVGGTVTINCQASENIYNWLAWYQQKP

575

GQPPKLLIYTVGDLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSSSYVDNV

SEQ ID NO: 347

METGLRWLLLVAVLKGVQCQEQLKESGGRLVTPGTPLTLTCTVSGFSLNDYAVGWFRQAPGKG

LEWIGYIRSSGTTAYATWAKGRFTISATSTTVDLKITSPTTEDTATYFCARGGAGSSGVWILD

GFAP

SEQ ID NO: 348

QASENIYNWLA

SEQ ID NO: 349

TVGDLAS

SEQ ID NO: 350

QQGYSSSYVDNV

SEQ ID NO: 351

DYAVG

SEQ ID NO: 352

YIRS SGTTAYATWAKG

SEQ ID NO: 353

GGAGSSGVWILDGFAP

SEQ ID NO: 354

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAAA

TGTGCCGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTC

ACCATCAATTGCCAGGCCAGTGAGAACATTTATAATTGGTTAGCCTGGTATCAGCAGAAACCA

GGGCAGCCTCCCAAGCTCCTGATCTATACTGTAGGCGATCTGGCATCTGGGGTCTCATCGCGG

TTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGAT

GCTGCCACTTACTATTGTCAACAGGGTTATAGTAGTAGTTATGTTGATAATGTT

SEQ ID NO: 355

576

INSTITUTE MtXX7.N0 of the? Ύ ·: αο

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTÓiCAG,ffeTCAGGAG cagctgaaggagtccgggggtcgcctggtcacgcctgggacacccutgacactcacctgcacA

GTCTCTGGATTCTCCCTCAATGACTATGCAGTGGGCTGGTTCCGCCAGGCTCCAGGGAAGGGG

CTGGAATGGATCGGATACATTCGTAGTAGTGGTACCACAGCCTACGCGACCTGGGCGAAAGGC

CGATTCACCATCTCCGCTACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAG

GACACGGCCACCTATTTCTGTGCCAGAGGGGGTGCTGGTAGTAGTGGTGTGTGGATCCTTGAT

GGTTTTGCTCCC

SEQ ID NO: 356

CAGGCCAGTGAGAACATTTATAATTGGTTAGCC

SEQ ID NO: 357

ACTGTAGGCGATCTGGCATCT

SEQ ID NO: 358

CAACAGGGTTATAGTAGTAGTTATGTTGATAATGTT

SEQ ID NO: 359

GACTATGCAGTGGGC

SEQ ID NO: 360

TACATTCGTAGTAGTGGTACCACAGCCTACGCGACCTGGGCGAAAGGC

SEQ ID NO: 361

GGGGGTGCTGGTAGTAGTGGTGTGTGGATCCTTGATGGTTTTGCTCCC

SEQ ID NO: 362

MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSSVSAAVGGTVTINCQASQSVYQNNYLSWFQQK

PGQPPKLLIYGAATLASGVPSRFKGSGSGTQFTLTISDLECDDAATYYCAGAYRDVDS

SEQ ID NO: 363

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPGASLTLTCTASGFSFTSTYYIYWVRQAPGKG

LEWIACIDAGSSGSTYYATWVNGRFTISKTSSTTVTLQMTSLTAADTATYFCAKWDYGGNVGW

577

MEXICAN INSTITUTE OF PROPERTY

INDUST-Tl / iL

GYDL

SEQ ID NO: 364

QASQSVYQNNYLS

SEQ ID NO: 365

GAATLAS

SEQ ID NO: 366

AGAYRDVDS

SEQ ID NO: 367

STYYIY

SEQ ID NO: 368

CIDAGSSGSTYYATWVNG

SEQ ID NO: 369

WDYGGNVGWGYDL

SEQ ID NO: 370

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCTCAAGTGCTGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGGCCAGTCAGAGTGTTTATCAGAACAACTACTTATCCTGGTTTCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCGGCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGAC

GATGCTGCCACTTACTACTGTGCAGGCGCTTATAGGGATGTGGATTCT

SEQ ID NO: 371

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

TTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGGGGCATCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCTTTACTAGTACCTACTACATCTACTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTGGAGTGGATCGCATGTATTGATGCTGGTAGTAGTGGTAGCACTTACTACGCGACCTGGGTG

578

INSTITUTE Μ £ λ: θ .. \ · Ω L £ LA FRGHLDAO

INDOS 1 KÍaL

AATGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTG

ACAGCCGCGGACACGGCCACCTATTTCTGTGCGAAATGGGATTATGGTGGTAATGTTGGTTGG

GGTTATGACTTG

SEQ ID NO: 372

CAGGCCAGTCAGAGTGTTTATCAGAACAACTACTTATCC

SEQ ID NO: 373

GGTGCGGCCACTCTGGCATCT

SEQ ID NO: 374

GCAGGCGCTTATAGGGATGTGGATTCT

SEQ ID NO: 375

AGTACCTACTACATCTAC

SEQ ID NO: 376

TGTATTGATGCTGGTAGTAGTGGTAGCACTTACTACGCGACCTGGGTGAATGGC

SEQ ID NO: 377

TGGGATTATGGTGGTAATGTTGGTTGGGGTTATGACTTG

SEQ ID NO: 378

MDTRAPTQLLGLLLLWLPGARCAFELTQTPSSVEAAVGGTVTIKCQASQSISSYLAWYQQKPG

QPPKFLIYRASTLASGVPSRFKGSGSGTEFTLTISDLECADAATYYCQSYYDSVSNP

SEQ ID NO: 379

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPEGSLTLTCKASGLDLGTYWFMCWVRQAPGKG

LEWIACIYTGSSGSTFYASWVNGRFTISKTSSTTVTLQMTSLTAADTATYFCARGYSGYGYFK

L

SEQ ID NO: 380

QASQSISSYLA

SEQ ID NO: 381

579

SEQ ID NO: 382 —-QSYYDSVSNP

SEQ ID NO: 383

TYWFMC

SEQ ID NO: 384

CIYTGSSGSTFYASWVNG

SEQ ID NO: 385

GYSGYGYFKL

SEQ ID NO: 386

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCATTCGAATTGACCCAGACTCCATCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTCAGAGCATTAGTAGTTACTTAGCCTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGTTCCTGATCTACAGGGCGTCCACTCTGGCATCTGGGGTCCCATCGCGATTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAAAGCTATTATGATAGTGTTTCAAATCCT

SEQ ID NO: 387

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

TTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGAGGGATCCCTGACACTCACCTGCAAAGCC

TCTGGACTCGACCTCGGTACCTACTGGTTCATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTGGAGTGGATCGCTTGTATTTATACTGGTAGTAGTGGTTCCACTTTCTACGCGAGCTGGGTG

AATGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTG

ACAGCCGCGGACACGGCCACTTATTTTTGTGCGAGAGGTTATAGTGGTTATGGTTATTTTAAG

TTG

SEQ ID NO: 388

580

CAGGCCAGTCAGAGCATTAGTAGTTACTTAGCC

ΙΜΡΪ

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

SEQ ID NO: 389 _-_ ~ -

AGGGCGTCCACTCTGGCATCT

SEQ ID NO: 390

CAAAGCTATTATGATAGTGTTTCAAATCCT

SEQ ID NO: 391

ACCTACTGGTTCATGTGC

SEQ ID NO: 392

TGTATTTATACTGGTAGTAGTGGTTCCACTTTCTACGCGAGCTGGGTGAATGGC

SEQ ID NO: 393

GGTTATAGTGGTTATGGTTATTTTAAGTTG

SEQ ID NO: 394

MDTRAPTQLLGLLLLWLPGVTFAIEMTQSPFSVSAAVGGTVSISCQASQSVYKNNQLSWYQQK

SGQPPKLLIYGASALASGVPSRFKGSGSGTEFTLTISDVQCDDAATYYCAGAITGSIDTDG

SEQ ID NO: 395

METGLRWLLLVAVLKGVQCQSLEESGGDLVKPGASLTLTCTTSGFSFSSSYFICWVRQAPGKG

LEWIACIYGGDGSTYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCAREWAYSQGYFG

AFDL

SEQ ID NO: 396

QASQSVYKNNQLS

SEQ ID NO: 397

FOOD

SEQ ID NO: 398,

AGAITGSIDTDG

SEQ ID NO: 399

581

PEE

INSTITUTO MEXICANO DELA i í.'OÍ'IEDA INDUSTRIAL

SSYFIC

SEQ ID NO: 400

CIYGGDGSTYYASWAKG

SEQ ID NO: 401

EWAYSQGYFGAFDL

SEQ ID NO: 402

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGTCACA

TTTGCCATCGAAATGACCCAGAGTCCATTCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGC

ATCAGTTGCCAGGCCAGTCAGAGTGTTTATAAGAACAACCAATTATCCTGGTATCAGCAGAAA

TCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCGGCTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTGCAGGCGCTATTACTGGTAGTATTGATACGGATGGT

SEQ ID NO: 403

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

TTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGGGGCATCCCTGACACTCACCTGCACAACT

TCTGGATTCTCCTTCAGTAGCAGCTACTTCATTTGCTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTGGAGTGGATCGCATGCATTTATGGTGGTGATGGCAGCACATACTACGCGAGCTGGGCGAAA

GGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACGCTGCAAATGACCAGTCTGACA

GCCGCGGACACGGCCACCTATTTCTGTGCGAGAGAATGGGCATATAGTCAAGGTTATTTTGGT

GCTTTTGATCTC

SEQ ID NO: 404

CAGGC CAGTCAGAGT GT TTATAAGAACAACCAATTATCC

SEQ ID NO: 405

GGTGCATCGGCTCTGGCATCT

SEQ ID NO: 406

582

IMPI

M £ X! CANO INSTITUTE

VAT AA and UTO MEXICANO J'i

.........

OF INDUSTRIAL PROPERTY

GCAGGCGCTATTACTGGTAGTATTGATACGGATGGT

SEQ ID NO: 407 '---- AGCAGCTACTTCATTTGC

SEQ ID NO: 408

TGCATTTATGGTGGTGATGGCAGCACATACTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 409

GAATGGGCATATAGTCAAGGTTATTTTGGTGCTTTTGATCTC

SEQ ID NO: 410

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASEDISSYLAWYQQKPG

QPPKLLIYAASNLESGVSSRFKGSGSGTEYTLTISDLECADAATYYCQCTYGTISISDGNA

SEQ ID NO: 411

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMTWVRQAPGEGL

EYIGFINPGGSAYYASWVKGRFTISKSSTTVDLKITSPTTEDTATYFCARVLIVSYGAFTI

SEQ ID NO: 412

QASEDISSYLA

SEQ ID NO: 413

AASNLES

SEQ ID NO: 414

QCTYGTISISDGNA

SEQ ID NO: 415

SYFMT

SEQ ID NO: 416

FINPGGSAYYASWVKG

SEQ ID NO: 417

VLIVSYGAFTI

583

IP

INSTITUTO MFALOPPj UE LA KROHELVU

INDUSTRIAL

SEQ ID NO: 418

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTGAGGATATTAGTAGCTACTTAGCCTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGCTCCTGATCTATGCTGCATCCAATCTGGAATCTGGGGTCTCATCGCGATTC

AAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACCTATTACTGTCAATGTACTTATGGTACTATTTCTATTAGTGATGGTAATGCT

SEQ ID NO: 419

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAATGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGATTCTCCCTCAGTAGCTACTTCATGACCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTG

GAATACATCGGATTCATTAATCCTGGTGGTAGCGCTTACTACGCGAGCTGGGTGAAAGGCCGA

TTCACCATCTCCAAGTCCTCGACCACGGTAGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGGGTTCTGATTGTTTCTTATGGAGCCTTTACCATC

SEQ ID NO: 420

CAGGCCAGTGAGGATATTAGTAGCTACTTAGCC

SEQ ID NO: 421

GCTGCATCCAATCTGGAATCT

SEQ ID NO: 422

CAATGTACTTATGGTACTATTTCTATTAGTGATGGTAATGCT

SEQ ID NO: 423

AGCTACTTCATGACC

SEQ ID NO: 424

TTCATTAATCCTGGTGGTAGCGCTTACTACGCGAGCTGGGTGAAAGGC

SEQ ID NO: 425

584

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

GTTCTGATTGTTTCTTATGGAGCCTTTACCATC

SEQ ID NO: 426

MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVSAAVGGTVTIKCQASEDIESYLAWYQQKPG

QPPKLLIYGASNLESGVSSRFKGSGSGTEFTLTISDLECADAATYYCQCTYGIISISDGNA

SEQ ID NO: 427

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMTWVRQAPGEGL

EYIGFMNTGDNAYYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARVLVVAYGAFNI

SEQ ID NO: 428

QASEDIESYLA

SEQ ID NO: 429

GASNLES

SEQ ID NO: 430

QCTYGIISISDGNA

SEQ ID NO: 431

SYFMT

SEQ ID NO: 432

FMNTGDNAYYASWAKG

SEQ ID NO: 433

VLVVAYGAFNI

SEQ ID NO: 434

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTGAGGACATTGAAAGCTATCTAGCCTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGAATCTGGGGTCTCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

585

M¿X INSTITUTE: O,

OF INDUSTRIAL PROPERTY

GCCACTTACTATTGTCAATGCACTTATGGTATTATTAGTATTAGTGATGGTAATGCT

SEQ ID NO: 435

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTG

TCTGGATTCTCCCTCAGTAGCTACTTCATGACCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTG

GAATACATCGGATTCATGAATACTGGTGATAACGCATACTACGCGAGCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGGGTTCTTGTTGTTGCTTATGGAGCCTTTAACATC

SEQ ID NO: 436

CAGGCCAGTGAGGACATTGAAAGCTATCTAGCC

SEQ ID NO: 437

GGTGCATCCAATCTGGAATCT

SEQ ID NO: 438

CAATGCACTTATGGTATTATTAGTATTAGTGATGGTAATGCT

SEQ ID NO: 439

AGCTACTTCATGACC

SEQ ID NO: 440

TTCATGAATACTGGTGATAACGCATACTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 441

GTTCTTGTTGTTGCTTATGGAGCCTTTAACATC

SEQ ID NO: 442

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSEPVGGTVSISCQSSKSVMNNNYLAWYQQK

PGQPPKLLIYGASNLASGVPSRFSGSGSGTQFTLTISDVQCDDAATYYCQGGYTGYSDHGT

SEQ ID NO: 443

METGLRWLLLVAVLKGVQCQSVEESGGRLVKPDETLTLTCTVSGIDLSSYPMNWVRQAPGKGL

586

EWIGFINTGGTIVYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARGSYVSSGYAYYFN

V

SEQ ID NO: 444

QSSKSVMNNNYLA

SEQ ID NO: 445

GASNLAS

SEQ ID NO: 446

QGGYTGYSDHGT

SEQ ID NO: 447

SYPMN

SEQ ID NO: 448

FINTGGTIVYASWAKG

SEQ ID NO: 449

GSYVSSGYAYYFNV

SEQ ID NO: 450

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGAACCTGTGGGAGGCACAGTCAGC

ATCAGTTGCCAGTCCAGTAAGAGTGTTATGAATAACAACTACTTAGCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGCATCTGGGGTCCCATCA

CGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGAC

GATGCTGCCACTTACTACTGTCAAGGCGGTTATACTGGTTATAGTGATCATGGGACT

SEQ ID NO: 451

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCAAGCCTGACGAAACCCTGACACTCACCTGCACAGTC

TCTGGAATCGACCTCAGTAGCTATCCAATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

587

IMPI

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

GAATGGATCGGATTCATTAATACTGGTGGTACCATAGTCTACGCGAGCTGGGCAAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGGCAGTTATGTTTCATCTGGTTATGCCTACTATTTTAAT

GTC

SEQ ID NO: 452

CAGTCCAGTAAGAGTGTTATGAATAACAACTACTTAGCC

SEQ ID NO: 453

GGTGCATCCAATCTGGCATCT

SEQ ID NO: 454

CAAGGCGGTTATACTGGTTATAGTGATCATGGGACT

SEQ ID NO: 455

AGCTATCCAATGAAC

SEQ ID NO: 456

TTCATTAATACTGGTGGTACCATAGTCTACGCGAGCTGGGCAAAAGGC

SEQ ID NO: 457

GGCAGTTATGTTTCATCTGGTTATGCCTACTATTTTAATGTC

SEQ ID NO: 458

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQSSQSVYNNNWLSWFQQK

PGQPPKLLIYKASTLASGVPSRFKGSGSGTQFTLTISDVQCDDVATYYCAGGYLDSVI

SEQ ID NO: 459

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSTYSINWVRQAPGKGL

EWIGIIANSGTTEYANWAKGRFTVSKTSTTVDLKITSPTTEDTATYFCARESGMYNEYGKFNI

SEQ ID NO: 460

QSSQSVYNNNWLS

SEQ ID NO: 461

588

IM F

KASTLAS

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

SEQ ID NO: 462

AGGYLDSVI

SEQ ID NO: 463

TYSIN

SEQ ID NO: 464

IIANSGTTFYANWAKG

SEQ ID NO: 465

ESGMYNEYGKFNI

SEQ ID NO: 466

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGC

ATCAGTTGCCAGTCCAGTCAGAGTGTTTATAATAACAACTGGTTATCCTGGTTTCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGAC

GATGTTGCCACTTACTACTGTGCGGGCGGTTATCTTGATAGTGTTATT

SEQ ID NO: 467

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGATTCTCCCTCAGTACCTATTCAATAAACTGGGTCCGCCAGGCTCCAGGGAAGGGCCTG

GAATGGATCGGAATCATTGCTAATAGTGGTACCACATTCTACGCGAACTGGGCGAAAGGCCGA

TTCACCGTCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGAGAGTGGAATGTACAATGAATATGGTAAATTTAACATC

SEQ ID NO: 468

CAGTCCAGTCAGAGTGTTTATAATAACAACTGGTTATCC

589

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

SEQ ID NO: 469

AAGGCATCCACTCTGGCATCT

SEQ ID NO: 470

GCGGGCGGTTATCTTGATAGTGTTATT

SEQ ID NO: 471

ACCTATTCAATAAAC

SEQ ID NO: 472

ATCATTGCTAATAGTGGTACCACATTCTACGCGAACTGGGCGAAAGGC

SEQ ID NO: 473

GAGAGTGGAATGTACAATGAATATGGTAAATTTAACATC

SEQ ID NO: 474

MDTRAPTQLLGLLLLWLPGARCASDMTQTPSSVSAAVGGTVTINCQASENIYSFLAWYQQKPG

QPPKLLIFKASTLASGVSSRFKGSGSGTQFTLTISDLECDDAATYYCQQGATVYDIDNN

SEQ ID NO: 475

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGIDLSAYAMIWVRQAPGEGL

EWITIIYPNGITYYANWAKGRFTVSKTSTAMDLKITSPTTEDTATYFCARDAESSKNAYWGYF

NV

SEQ ID NO: 476

QASENIYSFLA

SEQ ID NO: 477

KASTLAS

SEQ ID NO: 478

QQGATVYDIDNN

SEQ ID NO: 479

AYAMI

590

INÓilTU.O iVEXiC / .Nj D £ LA PKOl'iEDAD

INDUSTRIAL

SEQ ID NO: 480

IIYPNGITYYANWAKG

SEQ ID NO: 481

DAESSKNAYWGYFNV

SEQ ID NO: 482

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTCTGATATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGGCCAGTGAGAACATTTATAGCTTTTTGGCCTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGCTCCTGATCTTCAAGGCTTCCACTCTGGCATCTGGGGTCTCATCGCGGTTC

AAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCT

GCCACTTACTACTGTCAACAGGGTGCTACTGTGTATGATATTGATAATAAT

SEQ ID NO: 483

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTT

TCTGGAATCGACCTCAGTGCCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTG

GAATGGATCACAATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAAGGCCGA

TTCACCGTCTCCAAAACCTCGACCGCGATGGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGATGCAGAAAGTAGTAAGAATGCTTATTGGGGCTACTTT

AACGTC

SEQ ID NO: 484

CAGGCCAGTGAGAACATTTATAGCTTTTTGGCC

SEQ ID NO: 485

AAGGCTTCCACTCTGGCATCT

SEQ ID NO: 486

CAACAGGGTGCTACTGTGTATGATATTGATAATAAT

591 iMPia

Y'- 'J. MEXICAN INSTITUTE. TO

DELA PROPERTY V a-üíi-f __ INDUSTRIAL

SEQ ID NO: 487

GCCTATGCAATGATC -

SEQ ID NO: 488

ATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAAGGC

SEQ ID NO: 489

GATGCAGAAAGTAGTAAGAATGCTTATTGGGGCTACTTTAACGTC

SEQ ID NO: 490

MDTRAPTQLLGLLLLWLPGARCASDMTQTPSSVSAAVGGTVTINCQASENIYSFLAWYQQKPG

QPPKLLIFRASTLASGVSSRFKGSGSGTQFTLTISDLECDDAATYYCQQGATVYDIDNN

SEQ ID NO: 491

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGIDLSAYAMIWVRQAPGEGL

EWITIIYPNGITYYANWAKGRFTVSKTSTAMDLKITSPTTEDTATYFCARDAESSKNAYWGYF

NV

SEQ ID NO: 492

QASENIYSFLA

SEQ ID NO: 493

RASTLAS

SEQ ID NO: 494

QQGATVYDIDNN

SEQ ID NO: 495

AYAMI

SEQ ID NO: 496

11Y PNGIT YYANWAKG

SEQ ID NO: 497

DAESSKNAYWGYFNV

592

SEQ ID NO: 498

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTCTGATATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGGCCAGTGAGAACATTTATAGCTTTTTGGCCTGGTATCAGCAGAAACCAGGG

CAGCCTCCCAAGCTCCTGATCTTCAGGGCTTCCACTCTGGCATCTGGGGTCTCATCGCGGTTC

AAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCT

GCCACTTACTACTGTCAACAGGGTGCTACTGTGTATGATATTGATAATAAT

SEQ ID NO: 499

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTT

TCTGGAATCGACCTCAGTGCCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTG

GAATGGATCACAATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAAGGCCGA

TTCACCGTCTCCAAAACCTCGACCGCGATGGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGATGCAGAAAGTAGTAAGAATGCTTATTGGGGCTACTTT

AACGTC

SEQ ID NO: 500

CAGGCCAGTGAGAACATTTATAGCTTTTTGGCC

SEQ ID NO: 501

AGGGCTTCCACTCTGGCATCT

SEQ ID NO: 502

CAACAGGGTGCTACTGTGTATGATATTGATAATAAT

SEQ ID NO: 503

GCCTATGCAATGATC

SEQ ID NO: 504

ATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAAGGC

593

ΙΜΡ1(

1Ν5ΤΙ i 'J. JM ¿. · ¡TAÑO Y 'DEU? 5 ·.? ·> :? £ · ΛΟ í

SEQ ID NO: 505

GATGCAGAAAGTAGTAAGAATGCTTATTGGGGCTACTTTAACGTCSEQ ID NO: 506

MDTRAPTQLLGLLLLWLPGATFAIEMTQTPSPVSAAVGGTVTINCQASESVFNNMLSWYQQKP

GHSPKLLIYDASDLASGVPSRFKGSGSGTQFTLTISGVECDDAATYYCAGYKSDSNDGDNV

SEQ ID NO: 507

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLNRNSITWVRQAPGEGL

EWIGIITGSGRTYYANWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARGHPGLGSGNI

SEQ ID NO: 508

QASESVFNNMLS

SEQ ID NO: 509

DASDLAS

SEQ ID NO: 510

AGYKSDSNDGDNV

SEQ ID NO: 511

RNSIT

SEQ ID NO: 512

IITGSGRTYYANWAKG

SEQ ID NO: 513

GHPGLGSGNI

SEQ ID NO: 514

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCATTGAAATGACCCAGACTCCATCCCCCGTGTCTGCCGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGGCCAGTGAGAGTGTTTTTAATAATATGTTATCCTGGTATCAGCAGAAACCA

GGGCACTCTCCTAAGCTCCTGATCTATGATGCATCCGATCTGGCATCTGGGGTCCCATCGCGG

594

TTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGTGGCGTGGAGTGTGACGAT

GCTGCCACTTACTATTGTGCAGGGTATAAAAGTGATAGTAATGATGGCGATAATGTT

SEQ ID NO: 515

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGATTCTCCCTCAACAGGAATTCAATAACCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTG

GAATGGATCGGAATCATTACTGGTAGTGGTAGAACGTACTACGCGAACTGGGCAAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGGCCATCCTGGTCTTGGTAGTGGTAACATC

SEQ ID NO: 516

CAGGCCAGTGAGAGTGTTTTTAATAATATGTTATCC

SEQ ID NO: 517

GATGCATCCGATCTGGCATCT

SEQ ID NO: 518

GCAGGGTATAAAAGTGATAGTAATGATGGCGATAATGTT

SEQ ID NO: 519

AGGAATTCAATAACC

SEQ ID NO: 520

ATCATTACTGGTAGTGGTAGAACGTACTACGCGAACTGGGCAAAAGGC

SEQ ID NO: 521

GGCCATCCTGGTCTTGGTAGTGGTAACATC

SEQ ID NO: 522

MDTRAPTQLLGLLLLWLPGATFAQVLTQTASSVSAAVGGTVTINCQSSQSVYNNYLSWYQQKP

GQPPKLLIYTASSLASGVPSRFKGSGSGTQFTLTISEVQCDDAATYYCQGYYSGPIIT

SEQ ID NO: 523

595

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLNNYYIQWVRQAPGEGL

EWIGIIYAGGSAYYATWANGRFTIAKTSSTTVDLKMTSLTTEDTATYFCARGTFDGYEL

SEQ ID NO: 524

QSSQSVYNNYLS

SEQ ID NO: 525

TASSLAS

SEQ ID NO: 526

QGYYSGPIIT

SEQ ID NO: 527

NYYIQ

SEQ ID NO: 528

IIYAGGSAYYATWANG

SEQ ID NO: 529

GTFDGYEL

SEQ ID NO: 530

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCGCAAGTGCTGACCCAGACTGCATCGTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGTCCAGTCAGAGTGTTTATAATAACTACTTATCCTGGTATCAGCAGAAACCA

GGGCAGCCTCCCAAGCTCCTGATCTATACTGCATCCAGCCTGGCATCTGGGGTCCCATCGCGG

TTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGAAGTGCAGTGTGACGAT

GCTGCCACTTACTACTGTCAAGGCTATTATAGTGGTCCTATAATTACT

SEQ ID NO: 531

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAATAACTACTACATACAATGGGTCCGCCAGGCTCCAGGGGAGGGGCTG

596

MEXICAN INSTITUTE OE INDUSTRIAL PROPERTY

GAATGGATCGGGATCATTTATGCTGGTGGTAGCGCATACTACGCGACCTGGGCAAACGGCCGA

TTCACCATCGCCAAAACCTCGTCGACCACGGTGGATCTGAAGATGACCAGTCTGACAACCGAG

GACACGGCCACCTATTTCTGTGCCAGAGGGACATTTGATGGTTATGAGTTG

SEQ ID NO: 532

CAGTCCAGTCAGAGTGTTTATAATAACTACTTATCC

SEQ ID NO: 533

ACTGCATCCAGCCTGGCATCT

SEQ ID NO: 534

CAAGGCTATTATAGTGGTCCTATAATTACT

SEQ ID NO: 535

AACTACTACATACAA

SEQ ID NO: 536

ATCATTTATGCTGGTGGTAGCGCATACTACGCGACCTGGGCAAACGGC

SEQ ID NO: 537

GGGACATTTGATGGTTATGAGTTG

SEQ ID NO: 538

MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSPVSVPVGDTVTISCQSSESVYSNNLLSWYQQK

PGQPPKLLIYRASNLASGVPSRFKGSGSGTQFTLTISGAQCDDAATYYCQGYYSGVINS

SEQ ID NO: 539

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMSWVRQAPGEGL

EYIGFINPGGSAYYASWASGRLTISKTSTTVDLKITSPTTEDTATYFCARILIVSYGAFTI

SEQ ID NO: 540

QSSESVYSNNLLS

SEQ ID NO: 541

RASNLAS

597

ΙΜΡΪ

MEXICAN INSTITUTE OF INC'USTKiAL PROPERTY

SEQ ID NO: 542

QGYYSGVINS '........ ..........

SEQ ID NO: 543

SYFMS

SEQ ID NO: 544

FINPGGSAYYASWASG

SEQ ID NO: 545

ILIVSYGAFTI

SEQ ID NO: 546

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCCAAGTGCTGACCCAGACTCCATCCCCTGTGTCTGTCCCTGTGGGAGACACAGTCACC

ATCAGTTGCCAGTCCAGTGAGAGCGTTTATAGTAATAACCTCTTATCCTGGTATCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTACAGGGCATCCAATCTGGCATCTGGTGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGCACAGTGTGAC

GATGCTGCCACTTACTACTGTCAAGGCTATTATAGTGGTGTCATTAATAGT

SEQ ID NO: 547

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTG

TCTGGATTCTCCCTCAGTAGCTACTTCATGAGCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTG

GAATACATCGGATTCATTAATCCTGGTGGTAGCGCATACTACGCGAGCTGGGCGAGTGGCCGA

CTCACCATCTCCAAAACCTCGACCACGGTAGATCTGAAAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGGATTCTTATTGTTTCTTATGGAGCCTTTACCATC

SEQ ID NO: 54S

CAGTCCAGTGAGAGCGTTTATAGTAATAACCTCTTATCC

SEQ ID NO: 549

598

IMPI

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

AGGGCATCCAATCTGGCATCT

SEQ ID NO: 550

CAAGGCTATTATAGTGGTGTCATTAATAGT

SEQ ID NO: 551

AGCTACTTCATGAGC

SEQ ID NO: 552

TTCATTAATCCTGGTGGTAGCGCATACTACGCGAGCTGGGCGAGTGGC

SEQ ID NO: 553

ATTCTTATTGTTTCTTATGGAGCCTTTACCATC

SEQ ID NO: 554

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQATESIGNELSWYQQKPG

QAPKLLIYSASTLASGVPSRFKGSGSGTQFTLTITGVECDDAATYYCQQGYSSANIDNA

SEQ ID NO: 555

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSKYYMSWVRQAPEKGL

KYIGYIDSTTVNTYYATWARGRFTISKTSTTVDLKITSPTSEDTATYFCARGSTYFTDGGHRL DL |

SEQ ID NO: 556

QATESIGNELS

SEQ ID NO: 557

SASTLAS

SEQ ID NO: 558

QQGYSSANIDNA

SEQ ID NO: 559

KYYMS

SEQ ID NO: 560

599

IMPI

Mexican Institute of Industrial Property

YIDSTTVNTYYATWARG

SEQ ID NO: 561

GSTYFTDGGHRLDL

SEQ ID NO: 562

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCACTGAGAGCATTGGCAATGAGTTATCCTGGTATCAGCAGAAACCAGGG

CAGGCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTC

AAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACCGGCGTGGAGTGTGATGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTAGTGCTAATATTGATAATGCT

SEQ ID NO: 563

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACCGTC

TCTGGATTCTCCCTCAGTAAGTACTACATGAGCTGGGTCCGCCAGGCTCCAGAGAAGGGGCTG

AAATACATCGGATACATTGATAGTACTACTGTTAATACATACTACGCGACCTGGGCGAGAGGC

CGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAGATCACCAGTCCGACAAGTGAG

GACACGGCCACCTATTTCTGTGCCAGAGGAAGTACTTATTTTACTGATGGAGGCCATCGGTTG

GATCTC

SEQ ID NO: 564

CAGGCCACTGAGAGCATTGGCAATGAGTTATCC

SEQ ID NO: 565

TCTGCATCCACTCTGGCATCT

SEQ ID NO: 566

CAACAGGGTTATAGTAGTGCTAATATTGATAATGCT

SEQ ID NO: 567

600

AAGTACTACATGAGC

SEQ ID NO: 568 _

TACATTGATAGTACTACTGTTAATACATACTACGCGACCTGGGCGAGAGGC

SEQ ID NO: 569

GGAAGTACTTATTTTACTGATGGAGGCCATCGGTTGGATCTC

SEQ ID NO: 570

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQATESIGNELSWYQQKPG

QAPKLLIYSASTLASGVPSRFKGSGSGTQFTLTITGVECDDAATYYCQQGYSSANIDNA

SEQ ID NO: 571

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSTYNMGWVRQAPGKGL

EWIGSITIDGRTYYASWAKGRFTVSKSSTTVDLKMTSLTTGDTATYFCARILIVSYGAFTI

SEQ ID NO: 572

QATESIGNELS

SEQ ID NO: 573

SASTLAS

SEQ ID NO: 574

QQGYSSANIDNA

SEQ ID NO: 575

TYNMG

SEQ ID NO: 576

SITIDGRTYYASWAKG

SEQ ID NO: 577

ILIVSYGAFTI

SEQ ID NO: 578

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

601

TGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCACTGAGAGCATTGGCAATGAGTTATCCTGGTATCAGCAGAAACCAGGG

CAGGCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTC

AAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACCGGCGTGGAGTGTGATGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTAGTGCTAATATTGATAATGCT

SEQ ID NO: 579

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTAACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGATTCTCCCTCAGTACCTACAACATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAAGTATTACTATTGATGGTCGCACATACTACGCGAGCTGGGCGAAAGGCCGA

TTCACCGTCTCCAAAAGCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGGGGAC

ACGGCCACCTATTTCTGTGCCAGGATTCTTATTGTTTCTTATGGGGCCTTTACCATC

SEQ ID NO: 580

CAGGCCACTGAGAGCATTGGCAATGAGTTATCC

SEQ ID NO: 581

TCTGCATCCACTCTGGCATCT

SEQ ID NO: 582

CAACAGGGT TATAGTAGT GCTAATAT T GATAAT GCT

SEQ ID NO: 583

ACCTACAACATGGGC

SEQ ID NO: 584

AGTATTACTATTGATGGTCGCACATACTACGCGAGCTGGGCGAAAGGC

SEQ ID NO: 585

ATTCTTATTGTTTCTTATGGGGCCTTTACCATC

SEQ ID NO: 586

602

VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST

YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQ ID NO: 587

GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCC

TCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGAT

AACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACC

TACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCC

TGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

SEQ ID NO: 588

ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY

SLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLF

PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVL

TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV

KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL

HNHYTQKSLSLSPGK

SEQ ID NO: 589

GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGC

ACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAAC

TCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTAC

TCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAAC

GTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAA

ACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTC

CCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTG

GACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCAT

AATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTC

603

API

MEXICAN INSTITUTE OF THE PRGPiSD / O IN DUST HJ AL

ACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAEGCC

CTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTG

TACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTC

AAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAAC

TACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACC

GTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTG

CACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA

SEQ ID NO: 590

VPPGEDSKDVAAPHR

SEQ ID NO: 591

GEDSKDVAAPHRQPL

SEQ ID NO: 592

SKDVAAPHRQPLTSS

SEQ ID NO: 593

VAAPHRQPLTSSERI

SEQ ID NO: 594

PHRQPLTSSERIDKQ

SEQ ID NO: 595

QPLTSSERIDKQIRY

SEQ ID NO: 596

TSSERIDKQIRYILD

SEQ ID NO: 597

ERIDKQIRYILDGIS

SEQ ID NO: 598

DKQIRYILDGISALR

604

SEQ ID NO: 599

INSTITUTO MEXlC, DE LA FR0W £

INDUiT

IRYILDGISALRKET

SEQ ID NO: 600

ILDGISALRKETCNK

SEQ ID NO: 601

GISALRKETCNKSNM

SEQ ID NO: 602

ALRKETCNKSNMCES

SEQ ID NO: 603

KETCNKSNMCESSKE

SEQ ID NO: 604

CNKSNMCESSKEALA

SEQ ID NO: 605

SNMCESSKEALAENN

SEQ ID NO: 606

CESSKEALAENNLNL

SEQ ID NO: 607

SKEALAENNLNLPKM

SEQ ID NO: 608

ALAENNLNLPKMAEK

SEQ ID NO: 609

ENNLNLPKMAEKDGC

SEQ ID NO: 610

LNLPKMAEKDGCFQS

SEQ ID NO: 611

605

PKMAEKDGCFQSGFN

SEQ ID NO: 612

AEKDGCFQSGFNEET

SEQ ID NO: 613

DGCFQSGFNEETCLV

SEQ ID NO: 614

FQSGFNEETCLVKII

SEQ ID NO: 615

GFNEETCLVKIITGL

SEQ ID NO: 616

EETCLVKIITGLLEF

SEQ ID NO: 617

CLVKIITGLLEFEVY

SEQ ID NO: 618

KIITGLLEFEVYLEY

SEQ ID NO: 619

TGLLEFEVYLEYLQN

SEQ ID NO: 620

LEFEVYLEYLQNRFE

SEQ ID NO: 621

EVYLEYLQNRFESSE

SEQ ID NO: 622

LEYLQNRFESSEEQA

SEQ ID NO: 623

LQNRFESSEEQARAV

IMPI

INSTITUTO .MEXICANO D £ LA PROPIEDAD INDUSTRIAL

606

SEQ ID NO: 624

RFESSEEQARAVQMS

SEQ ID NO: 625

SSEEQARAVQMSTKV

SEQ ID NO: 626

EQARAVQMSTKVLIQ

SEQ ID NO: 627

RAVQMSTKVLIQFLQ

SEQ ID NO: 628

QMSTKVLIQFLQKKA

SEQ ID NO: 629

TKVLIQFLQKKAKNL

SEQ ID NO: 630

LIQFLQKKAKNLDAI

SEQ ID NO: 631

FLQKKAKNLDAITTP

SEQ ID NO: 632

KKAKNLDAITTPDPT

SEQ ID NO: 633

KNLDAITTPDPTTNA

SEQ ID NO: 634

DAITTPDPTTNASLL

SEQ ID NO: 635

TTPDPTTNASLLTKL

SEQ ID NO: 636

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

607

DPTTNASLLTKLQAQ

SEQ ID NO: 637 ~

TNASLLTKLQAQNQW

SEQ ID NO: 638

SLLTKLQAQNQWLQD

SEQ ID NO: 639

TKLQAQNQWLQDMTT

SEQ ID NO: 640

QAQNQWLQDMTTHLI

SEQ ID NO: 641

NQWLQDMTTHLILRS

SEQ ID NO: 642

LQDMTTHLILRSFKE

SEQ ID NO: 643

MTTHLILRSFKEFLQ

SEQ ID NO: 644

HLILRSFKEFLQSSL

SEQ ID NO: 645

LRSFKEFLQSSLRAL

SEQ ID NO: 646

FKEFLQSSLRALRQM

SEQ ID NO: 647

AYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKPGQRPKLLIYRASTLASGVSSRFK

GSGSGTE FTLTIS DLECADAAT AND YCQQGYSLRNIDNAFGGGTEVWKR

SEQ ID NO: 648

608

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

AIQMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQQKPGKAPKLLIYAASSLQSGVPSRFS

GSGSGTDFTLTISSLQPEDFATYYC

SEQ ID NO: 649

DIQMTQSPSSLSASVGDRVTITCRASQGISNYLAWYQQKPGKVPKLLIYAASTLQSGVPSRFS

GSGSGTDFTLTISSLQPEDVATYYC

SEQ ID NO: 650

DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLESGVPSRFS

GSGSGTEFTLTISSLQPDDFATYYC

SEQ ID NO: 651

AIQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFS

GSGSGTDFTLTISSLQPEDFATYYCQQGYSLRNIDNAFGGGTKVEIKR

SEQ ID NO: 652

QSLEESGGRLVTPGTPLTLTCTASGFSLSNYYVTWVRQAPGKGLEWIGIIYGSDETAYATWAI

GRFTIS KT S TTVDLKMT SLTAADTATY FCARDDS S DWDAKFNLWGQGTLVTVS S

SEQ ID NO: 653

EVQLVESGGGLVQPGGSLRLSCAASGFTVSSNYMSWVRQAPGKGLEWVSVIYSGGSTYYADSV

KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR

SEQ ID NO: 654

EVQLVESGGGLIQPGGSLRLSCAASGFTVSSNYMSWVRQAPGKGLEWVSVIYSGGSTYYADSV

KGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR

SEQ ID NO: 655

EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSVIYSGGSSTYYADS

VKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK

SEQ ID NO: 656

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATWA

609

ΜΡΙ

I Μ r

MEXICAN INSTITUTE, *

OF THE PKOi'IEEV.L · Oa INDUSTRIAL · ** 7Ü; ',

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSS

SEQ ID NO: 657

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATSA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSS

SEQ ID NO: 658

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYVTWVRQAPGKGL

EWIGIIYGSDETAYATSAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNLW

GQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVK

SEQ ID NO: 659

IIYGSDETAYATSAIG

SEQ ID NO: 660

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKPG

QRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNA

SEQ ID NO: 661

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYVTWVRQAPGKGL

EWIGIIYGSDETAYATWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNL

SEQ ID NO: 662

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAACCAGGG

CAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCT

SEQ ID NO: 663

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

610

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATCATTTATGGTAGTGATGAAACGGCCTACGCGACCTGGGCGATAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCCGCGGAC

ACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAACTTG

SEQ ID NO: 664

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATWA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSSASTKGP

SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSW

TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD

TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQD

WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ

KSLSLSPGK

SEQ ID NO: 665

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATSA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSSASTKGP

SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV

TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD

TLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQD

WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ

KSLSLSPGK

SEQ ID NO: 666

IQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFSG

611

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

SGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQ

LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE

KHKVYACEVTHQGLSSPVTKSFNRGEC

SEQ ID NO: 667

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTINCQASETIYSWLSWYQQKPG

QPPKLLIYQASDLASGVPSRFSGSGAGTEYTLTISGVQCDDAATYYCQQGYSGSNVDNV

SEQ ID NO: 668

METGLRWLLLVAVLKGVQCQEQLKESGGRLVTPGTPLTLTCTASGFSLNDHAMGWVRQAPGKG

LEYIGFINSGGSARYASWAEGRFTISRTSTTVDLKMTSLTTEDTATYFCVRGGAVWSIHSFDP

SEQ ID NO: 669

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACC

ATCAATTGCCAGGCCAGTGAGACCATTTACAGTTGGTTATCCTGGTATCAGCAGAAGCCAGGG

CAGCCTCCCAAGCTCCTGATCTACCAGGCATCCGATCTGGCATCTGGGGTCCCATCGCGATTC

AGCGGCAGTGGGGCTGGGACAGAGTACACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTGGTAGTAATGTTGATAATGTT

SEQ ID NO: 670

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGGAG

CAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTTACCTGCACA

GCCTCTGGATTCTCCCTCAATGACCATGCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTGGAATACATCGGATTCATTAATAGTGGTGGTAGCGCACGCTACGCGAGCTGGGCAGAAGGC

CGATTCACCATCTCCAGAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAG

GACACGGCCACCTATTTCTGTGTCAGAGGGGGTGCTGTTTGGAGTATTCATAGTTTTGATCCC

SEQ ID NO: 671

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQASQSVYDNNYLSWFQQK

612

MEXICAN INSTITUTE ÜE THE PROPERTY

INDUSTRIAL _

PGQPPKLLIYGASTLASGVPSRFVGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDNA SEQ ID NO: 672 '

METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSVYYMNWVRQAPGKGL

EWIGFITMSDNINYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARSRGWGTMGRLDL

SEQ ID NO: 673

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGC

ATCAGTTGCCAGGCCAGTCAGAGTGTTTATGACAACAACTACTTATCCTGGTTTCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCGTGGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACAGACGTGCAGTGTGAC

GATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATAATGCC

SEQ ID NO: 674

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACCCCTGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAGTGTCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGATTCATTACAATGAGTGATAATATAAATTACGCGAGCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTACAATGGGTCGGTTGGATCTC

SEQ ID NO: 675

MDTRAPTQLLGLLLLWLPGAICDPVLTQTPSPVSAPVGGTVSISCQASQSVYENNYLSWFQQK

PGQPPKLLIYGASTLDSGVPSRFKGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDDA

SEQ ID NO: 676

METGLRWLLLVAVLKGVQCQEQLKESGGGLVTPGGTLTLTCTASGFSLNAYYMNWVRQAPGKG

LEWIGFITLNNNVAYANWAKGRFTFSKTSTTVDLKMTSPTPEDTATYFCARSRGWGAMGRLDL

SEQ ID NO: 677

613

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCATA

TGTGACCCTGTGCTGACCCAGACTCCATCTCCCGTATCTGCACCTGTGGGAGGCACAGTCAGC

ATCAGTTGCCAGGCCAGTCAGAGTGTTTATGAGAACAACTATTTATCCTGGTTTCAGCAGAAA

CCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGATTCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATTACAGACGTGCAGTGTGAC

GATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATGATGCC

SEQ ID NO: 678

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGTCAGGAG

CAGCTGAAGGAGTCCGGAGGAGGCCTGGTAACGCCTGGAGGAACCCTGACACTCACCTGCACA

GCCTCTGGATTCTCCCTCAATGCCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGG

CTGGAATGGATCGGATTCATTACTCTGAATAATAATGTAGCTTACGCGAACTGGGCGAAAGGC

CGATTCACCTTCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACACCCGAG

GACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTGCAATGGGTCGGTTGGATCTC

SEQ ID NO: 679

MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSPVSAAVGGTVTINCQASQSVDDNNWLGWYQQK

RGQPPKYLIYSASTLASGVPSRFKGSGSGTQFTLTISDLECDDAATYYCAGGFSGNIFA

SEQ ID NO: 680

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGL

EWIGIIGGFGTTYYATWAKGRFTISKTSTTVDLRITSPTTEDTATYFCARGGPGNGGDI

SEQ ID NO: 681

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCCCAAGTGCTGACCCAGACTCCATCGCCTGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAACTGCCAGGCCAGTCAGAGTGTTGATGATAACAACTGGTTAGGCTGGTATCAGCAGAAA

CGAGGGCAGCCTCCCAAGTACCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCG

CGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGAC

614

SEQ ID NO: 682 ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTC

TCTGGCTTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGAAAGGGGCTG

GAGTGGATCGGAATCATTGGTGGTTTTGGTACCACATACTACGCGACCTGGGCGAAAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAGAATCACCAGTCCGACAACCGAGGAC

ACGGCCACCTATTTCTGTGCCAGAGGTGGTCCTGGTAATGGTGGTGACATC

SEQ ID NO: 683

MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSVPVGGTVTIKCQSSQSVYNNFLSWYQQKP

GQPPKLLIYQASKLASGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNA

SEQ ID NO: 684

METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGIDLSDYAMSWVRQAPGKGL

EWIGIIYAGSGSTWYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARDGYDDYGDFDRL

DL

SEQ ID NO: 685

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACA

TTTGCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGTACCTGTGGGAGGCACAGTCACC

ATCAAGTGCCAGTCCAGTCAGAGTGTTTATAATAATTTCTTATCGTGGTATCAGCAGAAACCA

GGGCAGCCTCCCAAGCTCCTGATCTACCAGGCATCCAAACTGGCATCTGGGGTCCCAGATAGG

TTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGAT

GCTGCCACTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCT

SEQ ID NO: 686

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCG

GTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACGCTCACCTGCACAGTC

615

IMPI

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

TCTGGAATCGACCTCAGTGACTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATCATTTATGCTGGTAGTGGTAGCACATGGTACGCGAGCTGGGCGAAAGGC

CGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAG

GACACGGCCACCTATTTCTGTGCCAGAGATGGATACGATGACTATGGTGATTTCGATCGATTG

GATCTC

SEQ ID NO: 687

MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKSG

QRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNA

SEQ ID NO: 688

METGLRWLLLVAVLSGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYMTWVRQAPGKGL

EWIGMIYGSDETAYANWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNL

SEQ ID NO: 689

ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGA

TGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACAGTCACC

ATCAAATGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAATCAGGG

CAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTC

AAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCT

GCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCT

SEQ ID NO: 690

ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCTCAGGTGTCCAGTGTCAGTCG

CTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCC

TCTGGATTCTCCCTCAGTAACTACTACATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTG

GAATGGATCGGAATGATTTATGGTAGTGATGAAACAGCCTACGCGAACTGGGCGATAGGCCGA

TTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCCGCGGAC

ACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAACTTG

616

MEXICAN INSTITUTE OF PROPERTY

INDUSTRIAL

SEQ ID NO: 691

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYMTWVRQAPGKGLEWVGMIYGSDETAYANWA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSSASTKGP

SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV

TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD

TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQD

WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ

KSLSLSPGK

SEQ ID NO: 692

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYMTWVRQAPGKGLEWVGMIYGSDETAYANSA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSSASTKGP

SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV

TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD

TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQD

WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ

KSLSLSPGK

SEQ ID NO: 693

DIQMTQSPSTLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFS

GSGSGTEFTLTISSLQPDDFATYYCQQGYSLRNIDNAFGGGTKVEIKRTVAAPSVFIFPPSDE

QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY

EKHKVYACEVTHQGLSS PVTKS FNRGEC

SEQ ID NO: 694

CAGGCCAGTCAGAGCATTAACAATGAGTTATCC

617

IMPI

SEQ ID NO: 695 industrial

CAACAGGGT TATAGTCT GAGGAACAT T GATAAT GCT * --- SEQ ID NO: 696

ATCATCTATGGTAGTGATGAAACCGCCTACGCTACCTCCGCTATAGGC

SEQ ID NO: 697

GAT GATAGTAGT GACTGGGATGCAAAGT T CAAC T TG

SEQ ID NO: 698

GCTATCCAGATGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATC

ACTTGCCAGGCCAGTCAGAGCATTAACAATGAGTTATCCTGGTATCAGCAGAAACCAGGGAAA

GCCCCTAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGC

GGCAGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCA

ACTTATTACTGCCAACAGGGTTATAGTCTGAGGAACATTGATAATGCTTTCGGCGGAGGGACC

AAGGTGGAAATCAAACGTACG

SEQ ID NO: 699

AIQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFS

GSGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNAFGGGTKVEIKRT

SEQ ID NO: 700

GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCC

TGTGCAGCCTCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGTCAGGCTCCAGGG

AAGGGGCTGGAGTGGGTCGGCATCATCTATGGTAGTGATGAAACCGCCTACGCTACCTCCGCT

ATAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGC

CTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGATGATAGTAGTGACTGGGATGCA

AAGTTCAACTTGTGGGGCjCAAGGGACCCTCGTCACCGTCTCGAGC

SEQ ID NO: 701

GCTATCCAGATGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATC

618

IMPI

ACTTGCCAGGCCAGTCAGAGCATTAACAATGAGTTATCCTGGTATCAGCAGAAACCAGGGAAA

GCCCCTAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGC

GGCAGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCA

ACTTATTACTGCCAACAGGGTTATAGTCTGAGGAACATTGATAATGCTTTCGGCGGAGGGACC

AAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAG

CAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCC

AAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAG

CAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTAC

GAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAG

AGCTTCAACAGGGGAGAGTGT

SEQ ID NO: 702

AIQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFS

GSGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNAFGGGTKVEIKRTVAAPSVFIFPPSDE

QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY

EKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQ ID NO: 703

GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCC

TGTGCAGCCTCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGTCAGGCTCCAGGG

AAGGGGCTGGAGTGGGTCGGCATCATCTATGGTAGTGATGAAACCGCCTACGCTACCTCCGCT

ATAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGC

CTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGATGATAGTAGTGACTGGGATGCA

AAGTTCAACTTGTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCA

TCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGC

CTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGC

GGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTG

619

MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY

ACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGC

AACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCG

TGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC

ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGAC

CCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCG

CGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGAC

TGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAG

AAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCC

CGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGC

GACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCC

GTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAG

AAGAGCCTCTCCCTGTCTCCGGGTAAA

SEQ ID NO: 704

EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATSA

IGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNLWGQGTLVTVSSASTKGP

SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV

TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD

TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQD

WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS

DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ

KSLSLSPGK

SEQ ID NO: 705

ATGAAGTGGGTAACCTTTATTTCCCTTCTGTTTCTCTTTAGCAGCGCTTATTCCGCTATCCAG

ATGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAG

620

IMPI

GCCAGTCAGAGCATTAACAATGAGTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAG

CTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGA

TCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTAC

TGCCAACAGGGTTATAGTCTGAGGAACATTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAA

ATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA

TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAG

TGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC

AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC

AAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC

AGGGGAGAGTGT

SEQ ID NO: 706

MKWVTFISLLFLFSSAYSAIQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPK

LLIYRASTLASGVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNAFGGGTKVE

IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS

KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQ ID NO: 707

ATGAAGTGGGTAACCTTTATTTCCCTTCTGTTTCTCTTTAGCAGCGCTTATTCCGAGGTGCAG

CTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCC

TCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGTCAGGCTCCAGGGAAGGGGCTG

GAGTGGGTCGGCATCATCTATGGTAGTGATGAAACCGCCTACGCTACCTCCGCTATAGGCCGA

TTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGCCTGAGAGCT

GAGGACACTGCTGTGTATTACTGTGCTAGAGATGATAGTAGTGACTGGGATGCAAAGTTCAAC

TTGTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTC

CCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAG

GACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCAC

621

IMPI

INSTV 'Ih or .MEXICANO D = LA PRCCLÜAD

INDUSTRIAL

ACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCC

TCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAG

GTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCA

CCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATG

ATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTC

AAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAG

CAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAAT

GGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATC

TCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAG

ATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCC

GTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGAC

TCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGG

AACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTC

TCCCTGTCTCCGGGTAAA

SEQ ID NO: 708

MKWVTFISLLFLFSSAYSEVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGL

EWVGIIYGSDETAYATSAIGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFN

LWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH

TFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPA

PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE

QYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE

MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG

NVFSCSVMHEALHNHYTQKSLSLSPGK

SEQ ID NO: 709

AIQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFS

622

IMPI

GSGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNAFGGGTKVEIKR SEQ ID NO: 710 ""

RASQGIRNDLG

SEQ ID NO: 711

RASQGISNYLA

SEQ ID NO: 712

RASQSISSWLA

SEQ ID NO: 713

AASSLQS

SEQ ID NO: 714

AASTLQS

SEQ ID NO: 715

KASSLES

SEQ ID NO: 716

SNYMS

SEQ ID NO: 717

VIYSGGSTYYADSVKG

SEQ ID NO: 718

VIYSGGSSTYYADSVKG

SEQ ID NO: 719

ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY

SLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLF

PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVL

TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLV

KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL

623

IMPI

HNHYTQKSLSLSPGK

SEQ ID NO: 720

ATCCAGATGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACT

TGCCAGGCCAGTCAGAGCATTAACAATGAGTTATCCTGGTATCAGCAGAAACCAGGGAAAGCC

CCTAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGC

AGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACT

TATTACTGCCAACAGGGTTATAGTCTGAGGAACATTGATAATGCT

SEQ ID NO: 721

GCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACAGTCACCATC

AAGTGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAACCAGGGCAG

CGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAA

GGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCC

ACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCTTTCGGCGGAGGGACC

GAGGTGGTGGTCAAACGT

SEQ ID NO: 722

ATCCAGATGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACT

TGCCAGGCCAGTCAGAGCATTAACAATGAGTTATCCTGGTATCAGCAGAAACCAGGGAAAGCC

CCTAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGC

AGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACT

TATTACTGCCAACAGGGTTATAGTCTGAGGAACATTGATAATGCTTTCGGCGGAGGGACCAAG

GTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAG

TTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAA

GTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAG

GACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAG

AAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGC

624

TTCAACAGGGGAGAGTGT

SEQ ID NO: 723

GCTATCCAGATGACCCAGTCTCCTTCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATC

ACTTGCCAGGCCAGTCAGAGCATTAACAATGAGTTATCCTGGTATCAGCAGAAACCAGGGAAA

GCCCCTAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGC

GGCAGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCA

ACTTATTACTGCCAACAGGGTTATAGTCTGAGGAACATTGATAATGCTTTCGGCGGAGGGACC

AAGGTGGAAATCAAACGT

SEQ ID NO: 724

GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCC

TGTGCAGCCTCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGTCAGGCTCCAGGG

AAGGGGCTGGAGTGGGTCGGCATCATCTATGGTAGTGATGAAACCGCCTACGCTACCTCCGCT

ATAGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACCCTGTATCTTCAAATGAACAGC

CTGAGAGCTGAGGACACTGCTGTGTATTACTGTGCTAGAGATGATAGTAGTGACTGGGATGCA

AAGTTCAACTTG

SEQ ID NO: 725

CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTCGGACACTCACCTGC

ACAGCCTCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGCCAGGCTCCAGGGAAG

GGGCTGGAATGGATCGGAATCATTTATGGTAGTGATGAAACGGCCTACGCGACCTGGGCGATA

GGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCC

GCGGACACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAAC

TTGTGGGGCCAAGGCACCCTGGTCACCGTCTCGAGC

SEQ ID NO: 726

MEKLLCFLVLTSLSHAFGQTDMSRKAFVFPKESDTSYVSLKAPLTKPLKAFTVCLHFYTELSS

TRGYSIFSYATKRQDNEILIFWSKDIGYSFTVGGSEILFEVPEVTVAPVHICTSWESASGIVE

625

IMPI

INSTITUTO MEXICANO DE UA FRO'f IEPáD fwvdgkprvrkslkkgytvgaeasiilgqeqdsfggnfegsqslvgdióWnmwdfvlspdei

NTIYLGGPFSPNVLNWRALKYEVQGEVFTKPQLWP

SEQ ID NO: 727

MLAVGCALLAALLAAPGAALAPRRCPAQEVARGVLTSLPGDSVTLTCPGVEPEDNATVHWVLR

KPAAGSHPSRWAGMGRRLLLRSVQLHDSGNYSCYRAGRPAGTVHLLVDVPPEEPQLSCFRKSP

LSNVVCEWGPRSTPSLTTKAVLLVRKFQNSPAEDFQEPCQYSQESQKFSCQLAVPEGDSSFYI

VSMCVASSVGSKFSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTWQDPHSWNSSFYRLR

FELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHWQLRAQEEFGQGEWSEWSPEAMGTPW

TESRSPPAENEVSTPMQALTTNKDDDNILFRDSANATSLPVQDSSSVPLPTFLVAGGSLAFGT

LLCIAIVLRFKKTWKLRALKEGKTSMHPPYSLGQLVPERPRPTPVLVPLISPPVSPSSLGSDN

TSSHNRPDARDPRSPYDISNTDYFFPR

SEQ ID NO: 728

MLTLQTWVVQALFIFLTTESTGELLDPCGYISPESPWQLHSNFTAVCVLKEKCMDYFHVNAN

YIVWKTNHFTIPKEQYTIINRTASSVTFTDIASLNIQLTCNILTFGQLEQNVYGITIISGLPP

EKPKNLSCIVNEGKKMRCEWDGGRETHLETNFTLKSEWATHKFADCKAKRDTPTSCTVDYSTV

YFVNIEVWVEAENALGKVTSDHINFDPVYKVKPNPPHNLSVINSEELSSILKLTWTNPSIKSV

IILKYNIQYRTKDASTWSQIPPEDTASTRSSFTVQDLKPFTEYVFRIRCMKEDGKGYWSDWSE

EASGITYEDRPSKAPSFWYKIDPSHTQGYRTVQLVWKTLPPFEANGKILDYEVTLTRWKSHLQ

NYTVNATKLTVNLTNDRYLATLTVRNLVGKSDAAVLTIPACDFQATHPVMDLKAFPKDNMLWV

EWTTPRESVKKYILEWCVLSDKAPCITDWQQEDGTVHRTYLRGNLAESKCYLITVTPVYADGP

GSPESIKAYLKQAPPSKGPTVRTKKVGKNEAVLEWDQLPVDVQNGFIRNYTIFYRTIIGNETA

VNVDSSHTEYTLSSLTSDTLYMVRMAAYTDEGGKDGPEFTFTTPKFAQGEIEAIVVPVCLAFL

LTTLLGVLFCFNKRDLIKKHIWPNVPDPSKSHIAQWSPHTPPRHNFNSKDQMYSDGNFTDVSV

VEIEANDKKPFPEDLKSLDLFKKEKINTEGHSSGIGGSSCMSSSRPSISSSDENESSQNTSST

VQYSTWHSGYRHQVPSVQVFSRSESTQPLLDSEERPEDLQLVDHVDGGDGILPRQQYFKQNC

626

ΡΙ

MEXICAN INSTITUTE OF ΕΛ PROPERTY

INDUSTRIAL _

SQHESSPDISHFERSKQVSSVNEEDFVRLKQQISDHISQSCGSGQMKMFQEVSAADAFG

GQVERFETVGMEAATDEGMPKSYLPQTVRQGGYMPQ

627

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408 members in 31 offices

This recordMember, by publication datePriority claim21 more offices are in the list below

Priority claims35

Priority claims
Document	Office	Kind	Date
11781108	United States of America	P	2008-11-25
11783908	United States of America	P	2008-11-25
11786108	United States of America	P	2008-11-25
61117811	United States of America	–	2008-11-25
61117839	United States of America	–	2008-11-25
61117861	United States of America	–	2008-11-25
12366567	United States of America	–	2009-02-05
36656709	United States of America	A	2009-02-05
12191615	United States of America	–	2009-02-24
12391717	United States of America	–	2009-02-24
19161509	United States of America	A	2009-02-24
39171709	United States of America	A	2009-02-24
12399156	United States of America	–	2009-03-06
39915609	United States of America	A	2009-03-06
12502581	United States of America	–	2009-07-14
50258109	United States of America	A	2009-07-14
2009006273	United States of America	W	2009-11-24
12191615	–	–	–
12366567	–	–	–
12391717	–	–	–
12399156	–	–	–
12502581	–	–	–
61117811	–	–	–
61117839	–	–	–
61117861	–	–	–
US0906273	–	–	–
US20080117811P	–	–	–
US20080117839P	–	–	–
US20080117861P	–	–	–
US20090191615	–	–	–
US20090366567	–	–	–
US20090391717	–	–	–
US20090399156	–	–	–
US20090502581	–	–	–
WO2009US06273	–	–	–

Members408

Family members, oldest first
Document	Office	Kind
US2007269868A1	United States of America	A1	2007-11-22
AU2007307324A1	Australia	A1	2008-04-17
CA2652392A1	Canada	A1	2008-04-17
WO2008045140A1	World Intellectual Property Organization (WIPO)	A1	2008-04-17
AU2008254578A1	Australia	A1	2008-11-27
AU2008254584A1	Australia	A1	2008-11-27
CA2688146A1	Canada	A1	2008-11-27
CA2688829A1	Canada	A1	2008-11-27
CA2993715A1	Canada	A1	2008-11-27
WO2008144753A2	World Intellectual Property Organization (WIPO)	A2	2008-11-27
WO2008144757A1	World Intellectual Property Organization (WIPO)	A1	2008-11-27
WO2008144763A2	World Intellectual Property Organization (WIPO)	A2	2008-11-27
TW200902550A	Taiwan Province of China	A	2009-01-16
TW200902720A	Taiwan Province of China	A	2009-01-16
US2009022659A1	United States of America	A1	2009-01-22
WO2008144763A3	World Intellectual Property Organization (WIPO)	A3	2009-01-22
US2009028784A1	United States of America	A1	2009-01-29
WO2008144753A3	World Intellectual Property Organization (WIPO)	A3	2009-01-29
EP2021463A1	European Patent Office (EPO)	A1	2009-02-11
NO20084835L	Norway	L	2009-02-16
TW200911826A	Taiwan Province of China	A	2009-03-16
US2009104187A1	United States of America	A1	2009-04-23
MX2008014692A	Mexico	A	2009-08-18
EP2021463A4	European Patent Office (EPO)	A4	2009-09-02
US2009238825A1	United States of America	A1	2009-09-24
JP2009537176A	Japan	A	2009-10-29
US2009291077A1	United States of America	A1	2009-11-26
US2009291082A1	United States of America	A1	2009-11-26
US2009291089A1	United States of America	A1	2009-11-26
US2009297436A1	United States of America	A1	2009-12-03
US2009297513A1	United States of America	A1	2009-12-03
CN101600793A	China	A	2009-12-09
NO20093387L	Norway	L	2010-01-15
NO20190984A1	Norway	A1	2010-01-15
MX2009012492A	Mexico	A	2010-01-20
MX2009012493A	Mexico	A	2010-01-20
NO20093386L	Norway	L	2010-02-18
KR20100028569A	Republic of Korea	A	2010-03-12
KR20100028571A	Republic of Korea	A	2010-03-12
EP2162469A1	European Patent Office (EPO)	A1	2010-03-17
EP2164514A2	European Patent Office (EPO)	A2	2010-03-24
US2010129357A1	United States of America	A1	2010-05-27
AU2009323002A1	Australia	A1	2010-06-10
AU2009323007A1	Australia	A1	2010-06-10
AU2009323008A1	Australia	A1	2010-06-10
AU2009323009A1	Australia	A1	2010-06-10
CA2744400A1	Canada	A1	2010-06-10
US2010143294A1	United States of America	A1	2010-06-10
WO2010065072A1	World Intellectual Property Organization (WIPO)	A1	2010-06-10
WO2010065077A2	World Intellectual Property Organization (WIPO)	A2	2010-06-10
WO2010065078A1	World Intellectual Property Organization (WIPO)	A1	2010-06-10
WO2010065079A2	World Intellectual Property Organization (WIPO)	A2	2010-06-10
IL202232D0	Israel	D0	2010-06-16
IL202238D0	Israel	D0	2010-06-16
US2010150829A1	United States of America	A1	2010-06-17
CN101754772A	China	A	2010-06-23
US2010158859A1	United States of America	A1	2010-06-24
EA200901533A1	Eurasian Patent Organization (EAPO)	A1	2010-06-30
TW201023888A	Taiwan Province of China	A	2010-07-01
TW201023889A	Taiwan Province of China	A	2010-07-01
TW201023890A	Taiwan Province of China	A	2010-07-01
WO2010065079A3	World Intellectual Property Organization (WIPO)	A3	2010-07-29
TW201028164A	Taiwan Province of China	A	2010-08-01
JP2010527615A	Japan	A	2010-08-19
JP2010528589A	Japan	A	2010-08-26
HK1138796A1	Hong Kong, China	A1	2010-09-03
WO2010065077A3	World Intellectual Property Organization (WIPO)	A3	2010-09-10
CN101868477A	China	A	2010-10-20
US2010290993A1	United States of America	A1	2010-11-18
WO2010065072A8	World Intellectual Property Organization (WIPO)	A8	2010-12-16
AR074227A1	Argentina	A1	2010-12-29
CO6241159A2	Colombia	A2	2011-01-20
US7906117B2	United States of America	B2	2011-03-15
US7935340B2	United States of America	B2	2011-05-03
WO2011066369A2	World Intellectual Property Organization (WIPO)	A2	2011-06-03
WO2011066371A2	World Intellectual Property Organization (WIPO)	A2	2011-06-03
WO2011066374A2	World Intellectual Property Organization (WIPO)	A2	2011-06-03
WO2011066378A2	World Intellectual Property Organization (WIPO)	A2	2011-06-03
MX2011005396A	Mexico	A	2011-06-09
MX2011005406A	Mexico	A	2011-06-16
MX2011005408A	Mexico	A	2011-06-16
MX2011005411A	Mexico	A	2011-06-16
SG171804A1	Singapore	A1	2011-07-28
IL212728D0	Israel	D0	2011-07-31
IL195285D0	Israel	D0	2011-08-01
KR20110091780A	Republic of Korea	A	2011-08-12
KR20110091781A	Republic of Korea	A	2011-08-12
EP2361095A1	European Patent Office (EPO)	A1	2011-08-31
EP2361096A1	European Patent Office (EPO)	A1	2011-08-31
US2011217303A1	United States of America	A1	2011-09-08
WO2011066369A3	World Intellectual Property Organization (WIPO)	A3	2011-09-22
EP2367570A2	European Patent Office (EPO)	A2	2011-09-28
CL2011001221A1	Chile	A1	2011-09-30
KR20110112307A	Republic of Korea	A	2011-10-12
KR20110112308A	Republic of Korea	A	2011-10-12
US2011250201A1	United States of America	A1	2011-10-13
EP2376126A2	European Patent Office (EPO)	A2	2011-10-19
WO2011066371A3	World Intellectual Property Organization (WIPO)	A3	2011-10-20
WO2011066374A3	World Intellectual Property Organization (WIPO)	A3	2011-10-20
NZ572807A	New Zealand	A	2011-10-28

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4 legal events, as the office reported them to INPADOC

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Events

Legal events, newest first
	Event	Code
2016-04-21	Grant or registrationFG	FG
2015-08-11	Transfer or rightsGB	GB
2012-09-14	Transfer or rightsGB	GB
2012-09-14	Transfer or rightsGB	GB