High concentration antibody formulations

[0001] Highly concentrated liquid antibody preparations are useful for delivering doses in a smaller volume. However, very concentrated protein preparations create a number of problems. One of the problems is the instability caused by the formation of solid particles. Another problem is the increased viscosity as a result of the numerous intermolecular interactions caused by the macromolecular nature of antibodies. High viscosity formulations are difficult to manufacture, inject into the syringe and inject. The use of force when handling such sticky preparations leads to excessive foaming, which can lead to denaturation and deactivation of active biological substances.

[0002] US Patent No. 6,685,322 and US Patent Application Publication Nos. 2006/0182740, 2007/0172479, 2008/0160014 disclose antibody preparations and methods for their preparation. None of these publications disclose the antibodies mentioned herein.

[0003] WO2009 / 079471 discloses a method of treating bone fractures with anti-sclerostin antibodies. WO02 / 30463 discloses preparations containing concentrated proteins of reduced viscosity. F. He et al. (Analytical Biochemistry (2010), 399: 141-143) disclosed a high-efficiency dynamic light scattering method for measuring the viscosity of concentrated protein solutions.

SUMMARY OF THE INVENTION [0004] The present disclosure is based on the discovery that the addition of calcium acetate at a low concentration, e.g. 5-10 mM, reduces the effective viscosity of formulations containing a high concentration of a selected anti-sclerostin antibody. In contrast, the same concentration of calcium acetate did not significantly reduce the viscosity of other antibody preparations. The preparation is a sterile liquid preparation comprising (a) an anti-sclerostin antibody at a concentration of at least 70 mg / ml, wherein said antibody comprises the amino acid sequences of SEQ ID NO: 86 and SEQ ID NO: 84; and (b) calcium acetate at a concentration ranging from 1 mM to 20 mM or from about 5 mM to about 10 mM, said formulation having an absolute viscosity of 10 cP or less. The absolute viscosity as described in this document,

[0005] The calcium salt is calcium acetate. Alternatively, in certain embodiments, the calcium salt is present at a concentration reducing the viscosity of the antibody formulation by at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36% 37% 38% 39% 40% 41% 42% 43% 44% 45% 46% 47% 48% 49% 50% 51% 52% 53 %, 54%, 55%, 56%, 57%, 58%, 59%, 60% or more compared to the same calcium-deficient antibody preparation.

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and (b) calcium acetate at a concentration ranging from about 5 mM to about 15 mM or from about 5 mM to about 10 mM, this formulation having an absolute viscosity of about 10 cP or less. Alternatively, in certain embodiments, the calcium acetate is present at a concentration reducing the viscosity of the antibody formulation by at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36% , 37%, 38%, 39%. 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56% , 57%, 58%, 59%, 60% or more compared to the same calcium acetate-free antibody preparation. calcium acetate is present in a concentration reducing the viscosity of the antibody preparation by at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22 % 23% 24% 25% 26% 27% 28% 29% 30% 31% 32% 33% 34% 35% 36% 37% 38% 39%. 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56% , 57%, 58%, 59%, 60% or more compared to the same calcium acetate-free antibody preparation. calcium acetate is present in a concentration reducing the viscosity of the antibody preparation by at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22 % 23% 24% 25% 26% 27% 28% 29% 30% 31% 32% 33% 34% 35% 36% 37% 38% 39%. 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56% , 57%, 58%, 59%, 60% or more compared to the same calcium acetate-free antibody preparation.

[0007] There is also provided a method for reducing the viscosity of a protein preparation, which method comprises adding calcium acetate in a concentration between 1 mM and 20 mM to a sclerostin immunoglobulin preparation, wherein the formulation contains immunoglobulin at a concentration of 70 mg / ml to 200 mg / ml. and the sclerostin immunoglobulin comprises the amino acid sequences of SEQ ID NO: 86 and SEQ ID NO: 84 and wherein the viscosity of the calcium acetate formulation is reduced compared to the viscosity of the antibody preparation without calcium acetate.

[0008] In another aspect, the formulation is sterile and has an absolute viscosity of about 10 cP or less and contains: (a) Ab-5 at a concentration of at least 70 mg / ml to about 200 mg / ml; (b) calcium acetate at a concentration ranging from about 1 mM to about 20 mM; and (c) a polyol, such as for example sucrose, in an amount in the range of about 1% w / v. up to about 12% w / v. In certain embodiments, the polyol is present in an amount in the range of about 4% to 10%. The immunoglobulin comprises the amino acid sequences of SEQ ID NO: 86 (Ab-5 heavy chain variable region) and / or SEQ ID NO: 84 (Ab-5 light chain variable region).

[0009] In another aspect, the formulation is sterile and has an absolute viscosity of about 10 cP or less and contains (a) Ab-5 at a concentration of at least 70 mg / ml to about 200 mg / ml; (b) calcium acetate at a concentration ranging from about 1 mM to about 20 mM; and (c) a polyol, such as for example sucrose, in an amount ranging from about 4% w / v. up to about 6% w / v.

[0010] In any of the preceding aspects, in some embodiments, the formulation further comprises (c) an acetate buffer, e.g. sodium acetate, at a concentration of about 5mM to about 15mM or about 5mM to about 10mM. In some embodiments, the total acetate concentration is from about 10mM to about 50mM or from about 20mM to about 40mM.

[0011] In another aspect of the disclosure, the preparation is sterile and if in liquid or reconstituted liquid form, comprises (a) an anti-sclerostin antibody at a concentration of about 70 mg / ml to about 200 mg / ml, wherein the antibody comprises a kit. six CDRs selected from the group consisting of SEQ ID NO: 1-5 (CDR Ab-A and Ab-1), 15-20 (CDR Ab-B), 25-30 (CDR Ab-C), 35-40 (CDR AbPZ) / 4572 / AGR

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D), 45-50 (CDR Ab-2), 55-60 (CDR Ab-3 and Ab-15), 73-78 (CDR Ab-4 and Ab-5), 91-96 (CDR Ab-6) , 101-106 (CDR Ab-7), 111-116 (CDR Ab-8), 121-126 (CDR Ab-9), 131-136 (CDR Ab-10), 141-146 (CDR Ab-11 and Ab-16), 159-164 (CDR Ab-12), 169-174 (CDR Ab-13 and Ab-14), 187-192 (CDR Ab-17 and Ab-18), 201-206 (CDR Ab- 19, Ab-20 and Ab-23), 225-229 (CDR Ab-21 and Ab-22) or 239-244 (CDR Ab-24); and (b) an acetate salt and / or an acetate buffer in a concentration ranging from about 10 mM to about 50 mM acetate or from about 20 mM to about 40 mM acetate, the formulation having an absolute viscosity of about 10 cP or less. In some embodiments, the acetate buffer comprises calcium acetate and / or sodium acetate. Alternatively, in certain embodiments, the acetate salt and / or buffer is present at a concentration that reduces the viscosity of the antibody formulation by at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33% , 34%, 35%, 36%, 37%, 38%, 39%. 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56% , 57%, 58%, 59%, 60% or more compared to the same antibody preparation without acetate salt and / or buffer.

[0012] In any of the preceding aspects, in some embodiments, the total concentration of ions (cations and anions) in the solution is from about 20mM to about 70mM or from about 30mM to about 60mM. In any of these embodiments, the overall osmolarity is less than about 400 mOsm / l or 350 mOsm / l, and is preferably close to an isotonic osmolarity, e.g., 250-350 mOsm / l. In some embodiments, the preparation is hypotonic. For example, in such embodiments, the osmolality of the formulation is less than about 250 mOsm / L. In other embodiments, the formulation is hypertonic. Thus, in such embodiments, the overall osmolality of the formulation is greater than about 350 mOsm / L.

[0013] In any of the preparations described herein, the anti-sclerostin antibody in the formulation may contain mature heavy and / or light chain variable regions of any Ab-A, Ab-B, Ab-C, Ab-D, Ab-2 antibody. , Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab12, Ab-15, Ab-16, Ab-17 , Ab-19, Ab-21, Ab-23 or Ab-24. Thus, in certain embodiments of the disclosure, the antibody comprises amino acid sequences: SEQ ID NO: 14 (Ab-1 heavy chain variable region) and / or SEQ ID NO: 12 (Ab-1 light chain variable region); or SEQ ID NO: 68 (Ab-15 heavy chain variable region) and / or SEQ ID NO: 66 (Ab15 light chain variable region); or SEQ ID NO: 86 (Ab-5 heavy chain variable region) and / or SEQ ID NO: 84 (Ab-5 light chain variable region); or SEQ ID NO: 154 (heavy chain variable region Ab-16) and / or SEQ ID NO: 152 (light chain variable region Ab-16); or SEQ ID NO: 182 (Ab-14 heavy chain variable region) and / or SEQ ID NO: 180 (Ab-14 light chain variable region); or SEQ ID NO: 208 (Ab-19 heavy chain variable region) and / or SEQ ID NO: 207 (light chain variable region of Ab19); or SEQ ID NO: 216 (Ab-20 heavy chain variable region) and / or SEQ ID NO: 214 (Ab-20 light chain variable region); or SEQ ID NO: 220 (Ab-23 heavy chain variable region) and / or SEQ ID NO: 218 (Ab-23 light chain variable region); or SEQ ID NO: 238 (Ab-22 heavy chain variable region) and / or SEQ ID NO: 236 (Ab-22 light chain variable region). In some embodiments of the disclosure, the antibody contains mature heavy and / or light chains of any Ab-A, Ab-B, Ab-C, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-Ab antibodies. 7, Ab-8, Ab-9, Ab-10, Ab-11, Ab12, Ab-15, Ab-16, Ab-19, Ab-23 or Ab-24. In certain embodiments of the disclosure, the antibody comprises amino acid sequences obtainable by expression in mammalian cells

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In a preferred embodiment of the present invention, a cDNA encoding a heavy and / or light chain or alternatively a heavy and / or light chain variable region of any of the Ab-A, Ab-B, Ab-C, Ab-D, Ab-2, Ab-3 antibodies is used. , Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-15, Ab-16, Ab-19, Ab -23 or Ab-24, as described herein.

For any of the preparations described herein, the anti-sclerostin antibody may comprise CDRs or mature heavy and light chain variable regions or mature heavy and light chains of any Ab-4 or Ab-5; Ab-13 or Ab-14; or Ab-19, Ab-20 or Ab-23. For any of the preparations described herein, in some embodiments, the antibody binds to sclerostin having the sequence SEQ ID NO: 1 with a K D of 10^-7 or less (lower numbers mean higher binding affinity).

[0015] For any of the preparations described herein, in some embodiments, the antibody is in a formulation at a concentration of at least 120 mg / ml or at least 140 mg / ml. In any of the preparations described herein, in certain embodiments, the absolute viscosity of the formulation is about 8 cP or less or about 6 cP or less. In alternative embodiments, the antibody in the formulation is present at a concentration of about 70 mg / ml to about 130 mg / ml, the formulation having an absolute viscosity of about 10 cps or less.

[0016] In some embodiments, any of the formulations described herein further comprises a polyol, such as sucrose, for example in an amount in the range of about 4% w / v. up to around 6%. In some embodiments, the formulation contains about 9% sucrose. In some embodiments, any of the formulations described herein include optionally other pharmaceutically acceptable excipients, e.g. salt, buffer, amino acid, stabilizer, polyol, other tonicity agent, surfactant, nailing agent, cryoprotective agent, lyophilization preservative, an antioxidant, a metal ion, a chelating agent and / or a preservative. In some embodiments, the formulation contains less than 0.05% by weight of surfactant.

[0017] For any of the preparations described herein, in certain embodiments, the formulation has a pH in the range of from about 4.5 to about 6 or from about 5 to about 6 or from about 5 to about 5.5. In some embodiments, the formulation has a pH of 5.2.

[0018] Also described herein are preparations for use in methods of treating any of the disorders associated with bone density reduction, including, but not limited to: achondroplasia, clavicular-cystadenostasis, chondroma, fibrous dysplasia, Gaucher disease, hypophosphatemic rickets, Marfan syndrome , multiple hereditary exostoses, neurofibromatosis, congenital bone fragility, osteopetrosis, osteopathilitis, atherosclerotic lesions, osteoarthritis, pyogenic osteoarthritis, periodontal disease, loss of bone tissue due to antiepileptic drugs, primary or secondary hyperparathyroidism, familial hyperparathyroidism syndrome , loss of bone tissue associated with lack of body mass, osteoporosis in men,post-menopausal bone loss, osteoarthritis, renal osteodystrophy, bone infiltration disorders, loss of bone-related bone tissue, sterile osteonecrosis of the jaw, juvenile Paget's disease, meloreostosis, metabolic bone diseases, mastocytosis, anemia / cell disease

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Sickle, loss of bone tissue associated with organ transplants, bone loss associated with kidney transplants, lichen lupus erythematosus, ankylosing spondylitis, epilepsy, ankylosing spondylitis, epilepsy, juvenile arthritis, thalassemia, mucopolysaccharidosis, disease Fabry, Turner syndrome, Down's syndrome, Klinefelter's syndrome, leprosy, Perthe's disease, idiopathic scoliosis of adolescents, starting with the childhood of multi-system inflammatory disease, Winchester syndrome, Menkes disease, Wilson's disease, ischemic bone disease (e.g. Legga-Calve-Perthes disease or local migrating osteoporosis), anemia, steroid induced conditions, glucocorticoid-induced bone loss,bone loss due to heparin, bone marrow disorders, scurvy, malnutrition, calcium deficiency, osteoporosis, osteopenia, alcoholism, chronic liver disease, postmenopausal condition, chronic inflammation, rheumatoid arthritis, inflammatory bowel disease, ulcerative colitis, colitis, Crohn's disease, menstrual disorders, amenorrhea, pregnancy, diabetes, hyperthyroidism, thyroid disorders, parathyroid disorder, Cushing's disease, acromegaly, hypogonadism, immobilization or non-use, sympathetic dystrophy, localized osteoporosis, osteomalacia, bone loss associated with replacement joints, bone loss associated with HIV, loss of bone tissue associated with the loss of growth hormone,bone loss associated with cystic fibrosis, bone loss associated with chemotherapy, tumor-induced bone loss, cancer-related bone loss, hormonal ablation bone loss, multiple myeloma, bone loss induced by medication, anorexia nervosa, loss-related disease facial bone mass associated with cranial bone disease associated with bone marrow depletion disease, cranial bone loss disease, bone loss due to aging, age-related facial bone loss, age-related cranial bone loss, age-related loss jaw bones, age-related cranial bone loss or loss of bone associated with being weightless.tumor-induced bone loss, cancer-related bone loss, hormonal ablative bone loss, multiple myeloma, loss of bone tissue caused by drugs, anorexia nervosa, bone loss-related facial bone disease associated with the disease of cranial bone loss associated with the disease bone loss of the jaw, cranial bone loss disease, loss of bone associated with aging, age-related facial bone loss, age-related cranial bone loss, age-related bone loss of the jaw, age-related cranial bone loss or bone loss associated with staying in a state of weightlessness.tumor-induced bone loss, cancer-related bone loss, hormonal ablative bone loss, multiple myeloma, loss of bone tissue caused by drugs, anorexia nervosa, bone loss-related facial bone disease associated with the disease of cranial bone loss associated with the disease bone loss of the jaw, cranial bone loss disease, loss of bone associated with aging, age-related facial bone loss, age-related cranial bone loss, age-related bone loss of the jaw, age-related cranial bone loss or bone loss associated with staying in a state of weightlessness.disease-related facial bone loss associated with cranial bone disease associated with bone marrow disease, cranial bone loss, bone loss due to aging, age-related facial bone loss, age-related cranial bone loss, age-related bone loss of the jaw, age-related cranial bone loss, or bone loss associated with being weightless.disease-related facial bone loss associated with cranial bone disease associated with bone marrow disease, cranial bone loss, bone loss due to aging, age-related facial bone loss, age-related cranial bone loss, age-related bone loss of the jaw, age-related cranial bone loss, or bone loss associated with being weightless.age-related cranial bone loss or bone loss associated with being in a state of weightlessness.age-related cranial bone loss or bone loss associated with being in a state of weightlessness.

[0019] The formulations described herein in some embodiments are useful for improving results in orthopedic procedures, dental procedures, implant surgery, articular replacement, bone grafts, bone plastic surgery and bone repair, such as healing of cracks, healing of non-growing parts, healing of delayed union and facial reconstruction. One or more formulations may be administered prior to, during and / or after surgery, replacement, transplantation, surgery or repair.

[0020] Such methods may include administering the formulation in a therapeutically effective amount, eg, an amount effective to improve bone density, and may further comprise administering a second therapeutic agent.

[0021] Also disclosed herein is a vial, kit or container, e.g. a pre-filled syringe or injection device, containing the preparation described herein, and optionally a label containing instructions for using the appropriate volume or amount of formulation necessary to achieve a dose of about 0.5-20. mg / kg or 0.5-10 mg / kg body weight of the patient.

[0022] It should be understood that although various embodiments in the description of the invention have been presented in various circumstances using the term "comprises", a related embodiment can also be described using the terms "consisting of" or "substantially consisting of". It should be

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EP 2 569 010 B1 is understood to refer to one or more, for example "immunoglobulin molecule" means one or more molecules of an immunoglobulin. As such, the terms singular, "one or more" and "at least one" may be used interchangeably herein.

[0023] It should also be understood that when providing a range of values, the described feature may have a single value falling in the range. For example, "a pH from about pH 4 to about pH 6" may include, but is not limited to, pH 4, 4.2, 4.6, 5.1, 5.5 etc. and any value between these values. In addition, the term & quot; pH from about pH 4 to about pH 6 & quot; should not be understood to mean that the pH of the preparation can vary by 2 pH units ranging from pH 4 to pH 6 during storage, but that the pH of the solution can be selected in this range of values, and the pH is buffered at about this pH. In some embodiments, when the term "about" is used, this indicates the number plus or minus 5%, 10%, 15% or more of said number. The actual intended variability can be determined from the context.

[0024] For any of the ranges given herein, range limitations are included in the scope. However, the description also includes the same ranges where the bottom and / or top limits are not in the range. For those skilled in the art, from the totality of this application, including drawings and detailed description, additional features and variants of the invention will become apparent, and all such features are intended to be aspects of the invention. Similarly, the features of the invention described herein can be combined again into additional embodiments, which are also intended to be aspects of the invention, regardless of whether or not a given combination of features is specifically mentioned above as an aspect or embodiment of the invention. In addition, only such restrictions, which are described herein as key to the invention should be perceived as such; variants of the invention without restrictions not described herein as critical are considered aspects of the invention.

DETAILED DESCRIPTION [0025] Formulations containing high concentrations of the antibody are described herein that contain calcium salts and / or acetate salts or buffers to reduce viscosity, as well as containers or kits containing these formulations.

I. Antibodies in the Preparation [0026] In some embodiments, the anti-sclerostin antibody in the formulation is present in a concentration of at least about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / ml, about 75 mg / ml, about 76 mg / ml, about 77 mg / ml, about 78 mg / ml, about 79 mg / ml, about 80 mg / ml, about 81 mg / ml, about 82 mg / ml, about 83 mg / ml, about 84 mg / ml, about 85 mg / ml, about 86 mg / ml, about 87 mg / ml, about 88 mg / ml, about 89 mg / ml, about 90 mg / ml, about 91 mg / ml, about 92 mg / ml, about 93 mg / ml, about 94 mg / ml, about 95 mg / ml, about 96 mg / ml, about 97 mg / ml, about 98 mg / ml, about 99 mg / ml, about 100 mg / ml, about 101 mg / ml, about 102 mg / ml, about 103 mg / ml, about 104 mg / ml, about 105 mg / ml, about 106 mg / ml, about 107 mg / ml, about 108 mg / ml, about 109 mg / ml, about 110 mg / ml, about 111 mg / ml, about 112 mg / ml,about 113 mg / ml, about 114 mg / ml, about 115 mg / ml, about 116 mg / ml, about 117 mg / ml, about 118 mg / ml, about 119 mg / ml, about 120 mg / ml, about 121 mg / ml, about 122 mg / ml, about 123 mg / ml, about 124 mg / ml, about 125 mg / ml, about 126 mg / ml, about 127 mg / ml,

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about 180 mg / ml or about 170 mg / ml. Any range including a combination of the above restrictions is contemplated, including, but not limited to, from about 70 mg / mL to about 250 mg / mL, about 70 mg / mL to about 200 mg / mL, about 70 mg / mL to about 160 mg / ml, from about 100 mg / ml to about 250 mg / ml, from about 100 mg / l to about 200 mg / ml, or from about 100 mg / ml to about 180 mg / ml.

[0027] Antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab- 8, Ab-9, Ab10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab- 21, Ab-22, Ab-23 and Ab-24 have been previously described in US patent application No. 2007/0110747.

[0028] Sclerostin antibodies described herein bind to sclerostin having the sequence SEQ ID NO: 1 with a KD of 10^-6 or less or 10^-7 or less or 10^-8 or less or 10^-9 or less (lower numbers mean higher binding affinity). Affinity can be determined by any of the methods known in the art, including using Biacore technology.

[0029] In embodiments, the antibody comprises a heavy and / or light Ab-5 antibody chain. Full-length mature amino-acid amino acid sequences for antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-15, Ab-16, Ab-17, Ab-19, Ab-23 and Ab-24, including the constant region, are determined, respectively, in SEQ ID NOs: 8, 22, 32, 42, 52, 62, 80, 88, 98, 108, 118, 128, 138, 148, 166, 176, 184, 70, 210 222 and 246. Amino acid sequences full-length mature heavy chain for antibodies Ab-A, Ab-B, Ab-C, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab- 8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-15, Ab-16, Ab-19, Ab-23 and Ab24, including the constant region are defined in SEQ ID NO: 10, 24, 34, 44, 54, 64, 82, 90, 100, 110, 120, 130, 140, 150, 168, 178, 186, 72, 224 and 248.

[0030] Suitable cDNA sequences coding for the full-length light chain for antibodies Ab-A, AbB, Ab-C, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab- 7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-15, Ab-16, Ab19, Ab-23 and Ab-24, including the constant region are determined in SEQ ID, respectively. NO: 7, 21, 31, 41, 51, 61, 79, 87, 97, 107, 117, 127, 137, 147, 165, 175, 183, 69, 209, 221 and 245. Suitable cDNA sequences coding for the heavy chain of full length including the constant region for antibodies Ab-A, Ab-B, AbC, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8 , Ab-9, Ab-10, Ab-11, Ab-12, Ab-15, Ab-16, Ab-19, Ab23 and Ab-24 are identified in SEQ ID NOs: 9, 23, 33, 43, respectively. , 53, 63, 81, 89, 99, 109, 119, 129, 139, 149, 167, 177, 185, 71, 211, 223, and 247.

[0031] This antibody comprises the variable region of the heavy and / or light chain of the Ab-5 antibody. This antibody contains SEQ ID NO: 86 (Ab-5 heavy chain variable region) and SEQ ID NO: 84 (Ab-5 light chain variable region).

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[0032] In certain embodiments of the disclosure, the antibody comprises the CDRs set forth in SEQ ID NO: 1-5 (CDRs of Ab-A and Ab-1) or 15-20 (CDRs of Ab-B) or 25-30. (CDR Ab-C) or 35-40 (CDR Ab-D) or 45-50 (CDR Ab-2) or 55-60 (CDR Ab-3 and Ab-15) or 73-78 (CDR Ab-4 and Ab-5) or 91-96 (CDR Ab-6) or 101-106 (CDR Ab-7) or 111-116 (CDR Ab-8) or 121-126 (CDR Ab-9) or 131-136 (CDR Ab-10) or 141-146 (CDR Ab-11 and Ab-16) or 159-164 (CDR Ab-12) or 169-174 (CDR Ab-13 and Ab-14) or 187-192 (CDR Ab- 17 and Ab-18) or 201-206 (CDR Ab-19, Ab-20 and Ab-23) or 225-229 (CDR Ab-21 and Ab-22) or 239244 (CDR Ab-24).

[0033] In some embodiments, the antibody comprises amino acid sequences obtainable by expression in mammalian host cells of the heavy and / or light heavy or light chain cDNA or alternatively the heavy chain and / or light chain variable region of the Ab-5 antibody, as described herein. . In any of the preparations described herein, in some embodiments, the antibody is a tetrameric immunoglobulin consisting of two heavy chains and two light chains.

[0034] In certain embodiments of the disclosure, the antibody comprises a CDR of any Ab-A, Ab-B, Ab-C, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6 antibody. , Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab12, Ab-15, Ab-16, Ab-19, Ab-23 or Ab-24 and contains a heavy and / or light chain containing an amino acid sequence of at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to the heavy and / or light chain of the antibody Ab-A, Ab- B, Ab-C, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-15, Ab-16, Ab-19, Ab-23 or Ab-24. In certain embodiments of the disclosure, the antibody comprises a CDR of any of Ab-A, Ab-B, AbC, Ab-D, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7 antibodies, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-15, Ab-16, Ab-19,

[0035] In certain embodiments of the disclosure, the antibody:

1) retains any one, two, three, four, five or six of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and / or CDRL3 of any Ab-A, Ab-B, Ab-C, Ab-D, Ab-Ab antibody 1, Ab-2, Ab-3, Ab4, Ab-5, Ab-6, Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab- 14, Ab-15, Ab-16, Ab-17, Ab-18, Ab19, Ab-20, Ab-21, Ab-22, Ab-23 or Ab-24 optionally including one or two mutations in such CDRs,

2) retains all of CDRH1, CDRH2, CDRH3 or heavy chain variable regions of any Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4 antibody, Ab-5, Ab-6, Ab7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab-21, Ab-22, Ab-23 or Ab-24 optionally including one or two mutations in such CDRs,

3) retains all of CDRL1, CDRL2, CDRL3 or light chain variable regions of any Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4 antibody, Ab-5, Ab-6, Ab-7, Ab-8,

Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab- 21, Ab-22,

Ab-23 or Ab-24 optionally including one or two mutations in such CDRs,

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4) binds to the same sclerostin epitope as the antibody Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6 , Ab-7, Ab-8, Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab -19, Ab-20, Ab-21, Ab-22, Ab-23 or Ab-24, e.g. as determined by X-ray crystallography or with the amino acid belonging to the loop formed by amino acids 86-111 of SEQ ID NO: 249; and / or

5) competes with the antibody Ab-A, Ab-B, Ab-C, Ab-D, Ab-1, Ab-2, Ab-3, Ab-4, Ab-5, Ab-6, Ab-7, Ab8 , Ab-9, Ab-10, Ab-11, Ab-12, Ab-13, Ab-14, Ab-15, Ab-16, Ab-17, Ab-18, Ab-19, Ab-20, Ab -21, Ab22, Ab-23 or Ab-24 in a binding to sclerostin greater than about 75%, greater than about 80% or greater than about 81%, 82%, 83%, 84%, 85%, 86% , 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% or 95%.

[0036] In some embodiments, the antibody comprises all three light chain CDRs, a mature light chain variable region, all three heavy chain CDRs, a mature heavy chain variable region, all six CDRs, or both a mature light chain variable region and a mature one. heavy chain variable region. In some exemplary embodiments of the disclosure, two CDRs of the light chain of one antibody can be combined with a third CDR of a light chain of another antibody. Alternatively, the CDRL1 of one antibody may be combined with the CDRL2 of another antibody and CDRL3 of yet another antibody, particularly when these CDRs are highly homologous. Similarly, two CDRs of the heavy chain of one antibody can be linked to the third CDR of the heavy chain of another antibody;

[0037] The term "antibody" refers to an intact antibody or a binding fragment thereof. The antibody can be a full antibody molecule (including polyclonal, monoclonal, chimeric, humanized or human versions containing heavy and / or full-length heavy chains) or an antigen-binding fragment thereof. Antibody fragments include F (ab ') 2, Fab, Fab', Fv, Fc and Fd fragments and can be incorporated into single domain antibodies, single chain antibodies, maxima, minibodies, intracells, diabodies, triabodies, tetrabodies, v-NARs and bis-scFv (see, e.g., Hollinger and Hudson, Nature Biotechnology, 23 (9): 1126-1136 (2005)).

[0038] The term "isolated" antibody refers to an antibody as defined herein that has been identified and separated from a component of its natural environment. The constituents of impurities from its natural environment are materials that could interfere with the diagnosis or therapeutic use of the antibody, and these may include enzymes, hormones and other proteinaceous or non-proteinaceous solutes. In certain embodiments, the antibody will be purified (1) to greater than 95% by weight of the antibody, and most preferably up to greater than 99% by weight, (2) sufficient to yield at least 15 N-terminal or internal amino acid residue or (3) homogeneity residues. in the SDS-PAGE assay under reducing or non-reducing conditions using Coomassie blue staining or, preferably, silver. An isolated naturally occurring antibody includes an antibody located in situ within recombinant cells, as it is the absence of at least one component of the antibody's natural environment. Typically, however, an isolated antibody will be produced using at least one purification step.

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[0039] An "immunoglobulin" or "native antibody" is a tetrameric glycoprotein. In a naturally occurring immunoglobulin, each tetramer consists of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kDa) and one "heavy" (about 50-70 kDa) chain. The portion of each amino terminated chain includes a "variable" ("V") region of from about 100 to about 110 or more amino acids primarily responsible for antigen recognition. The portion of each carboxyl terminated chain defines a constant region primarily responsible for effector function. Immunoglobulins can be assigned to different classes depending on the amino acid sequence of the constant domain of their heavy chains. Heavy chains are classified as mi (μ), delta (Δ), gamma (γ), alpha (α) and epsilon (ε) and they define the antibody's isotype as IgM, IgD, IgG, IgA and IgE, respectively. Many of these can be further subdivided into subclasses or isotypes, e.g. IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. Different isotypes have different effector functions; for example, IgG1 and IgG3 isotypes show antibody-dependent cellular cytotoxicity (ADCC) activity. Human light chains are classified as kappa (κ) and lambda (λ) light chains. In light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also containing the "D" region of about 10 consecutive amino acids. See generally Fundamental Immunology, ch. 7 (Paul, W., ed., Second edition, Raven Press, New York (1989)). Many of these can be further subdivided into subclasses or isotypes, e.g. IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. Different isotypes have different effector functions; for example, IgG1 and IgG3 isotypes show antibody-dependent cellular cytotoxicity (ADCC) activity. Human light chains are classified as kappa (κ) and lambda (λ) light chains. In light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also containing the "D" region of about 10 consecutive amino acids. See generally Fundamental Immunology, ch. 7 (Paul, W., ed., Second edition, Raven Press, New York (1989)). Many of these can be further subdivided into subclasses or isotypes, e.g. IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. Different isotypes have different effector functions; for example, IgG1 and IgG3 isotypes show antibody-dependent cellular cytotoxicity (ADCC) activity. Human light chains are classified as kappa (κ) and lambda (λ) light chains. In light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also containing the "D" region of about 10 consecutive amino acids. See generally Fundamental Immunology, ch. 7 (Paul, W., ed., Second edition, Raven Press, New York (1989)). for example, IgG1 and IgG3 isotypes show antibody-dependent cellular cytotoxicity (ADCC) activity. Human light chains are classified as kappa (κ) and lambda (λ) light chains. In light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also containing the "D" region of about 10 consecutive amino acids. See generally Fundamental Immunology, ch. 7 (Paul, W., ed., Second edition, Raven Press, New York (1989)). for example, IgG1 and IgG3 isotypes show antibody-dependent cellular cytotoxicity (ADCC) activity. Human light chains are classified as kappa (κ) and lambda (λ) light chains. In light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with the heavy chain also containing the "D" region of about 10 consecutive amino acids. See generally Fundamental Immunology, ch. 7 (Paul, W., ed., Second edition, Raven Press, New York (1989)). wherein the heavy chain also has a "D" region of about 10 consecutive amino acids. See generally Fundamental Immunology, ch. 7 (Paul, W., ed., Second edition, Raven Press, New York (1989)). wherein the heavy chain also has a "D" region of about 10 consecutive amino acids. See generally Fundamental Immunology, ch. 7 (Paul, W., ed., Second edition, Raven Press, New York (1989)).

[0040] Allotypes define antibody sequence variations, often in the constant region, that can be immunogenic and are encoded in humans by specific alleles. Allotypes for five of the IGHC human genes have been identified, namely IGHG1, IGHG2, IGHG3, IGHA2 and IGHE, and are designated as G1m, G2m, G3m, A2m and Em allotypes, respectively. At least 18 Gm allotypes are known: nG1m (1), nG1m (2), G1m (1, 2, 3, 17) or G1m (a, x, f, z), G2m (23) or G2m (n), G3m (5, 6, 10, 11, 13, 14, 15, 16, 21, 24, 26, 27, 28) or G3m (b1, c3, b5, b0, b3, b4, s, t, g1, c5, u, v, g5). There are two allotypes A2m: A2m (1) and A2m (2).

[0041] The term "hypervariable region" refers to amino acid residues from the complementarity determining region, CDRs (i.e. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31 -35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy-chain variable domain as described by Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, Maryland (1991)). Even a single CDR can recognize and bind antigen, although with lower affinity than the entire antigen binding site containing all CDRs.

[0042] An alternative definition of residues from the hypervariable "loop" is described by Chothia et al., J. Mol. Biol. 196: 901917 (1987) as residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2) and 96- 101 (H3) in the heavy chain variable domain.

[0043] "Skeletal" residues, FRs, are variable region residues other than hypervariable region residues.

[0044] "Antibody fragments" include a portion of an intact immunoglobulin, preferably an antigen binding or variable region of an intact antibody, and also include multispecific (bispecific, trispecific, etc.) antibodies formed from antibody fragments. Immunoglobulin fragments can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies.

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[0045] Non-limiting examples of antibody fragments include Fab, Fab ', F (ab') 2, Fv (variable region), domain antibodies (dAb, containing the VH domain) (Ward et al., Nature 341: 544 -546, 1989), fragments of the complementarity determining region (CDR), single-chain antibodies (scFv, containing the VH and VL domains on a single polypeptide chain) (Bird et al., Science

242: 423-426, 1988 and Huston et al., Prod. Natl. Acad. Sci. USA 85: 5879-5883, 1988, optionally including a polypeptide linkage and optionally multispecific linker, Gruber et al., J. Immunol. 152: 5368 (1994)), single chain antibody fragments, diabodies (VH and VL domains on a single polypeptide chain that bind to the complementary VL and VH domains of another chain) (EP 404097, WO 93/11161 and Hollinger et al., Proc Natl Acad Sci. USA, 90: 6444-6448 (1993)), triabodies, tetrabodies, minibodies (scFv fused to CH3 via a peptide linker (no hinge) or via IgG hinge) (Olafsen et al., Protein Eng Des Sel April 2004, 17 (4): 315-23), linear antibodies (tandem Fd segments (VH-CH1-VH-CH1) (Zapata et al., Protein Eng., 8 (10): 1057-1062 (1995) ) chelating recombinant antibodies (crAb,

[0046] The term "variant" when used in reference to antibodies refers to an antibody polypeptide sequence that comprises at least one amino acid substitution, deletion or insertion in a variable region or portion equivalent to a variable region, provided that the variant retains the desired binding affinity or activity. bioavailability. In addition, the antibodies described herein may comprise amino acid modifications in the constant region to modify the effector function of the antibody including the half-life or clearance, ADCC activity and / or CDC activity. Such modifications may, for example, improve the pharmacokinetics or increase the efficacy of the antibody in the treatment of cancer. See Shields et al., J. Biol. Chem., 276 (9): 6591-6604 (2001). In the case of IgG1, modifications of the constant region, particularly the hinge or CH2 region, they can increase or decrease effector function, including ADCC and / or CDC activity. The constant IgG2 region

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EP 2 569 010 B1 can be modified to reduce the formation of antibody-antigen aggregates. In the case of IgG4, modifications of the constant region, in particular the hinge region, may reduce the formation of half-antibodies.

[0047] The term "modification" when used in connection with the antibodies or polypeptides described herein includes, but is not limited to, one or more amino acid changes (including substitutions, insertions or deletions); chemical modifications that do not interfere with hepcidin binding activity; covalent modification by conjugation with therapeutic or diagnostic factors; labeling (e.g., radionuclides or various enzymes); covalent attachment of a polymer, such as pegylation (poly (ethylene glycol) derivation) and insertion or substitution of non-naturally occurring amino acids by chemical synthesis methods. Modified polypeptides (including antibodies) may retain the binding properties of the unmodified molecules of the invention.

The term "derivative" when used in connection with the antibodies or polypeptides of the invention refers to antibodies or polypeptides that have been covalently modified by conjugation with therapeutic or diagnostic agents, labeling (e.g., radionuclides or various enzymes), covalent attachment of a polymer such as pegylation (poly (ethylene glycol) derivation) and insertion or substitution of non-naturally occurring amino acids by chemical synthesis methods. Derivatives may retain the binding properties of non-derivatized molecules.

[0049] Methods for producing bispecific or other multispecific antibodies are known in the art and include chemical cross-linking, the use of leucine zippers [Kostelny et al., J. Immunol. 148: 1547-1553, 1992]; dialectal technology [Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-48, 1993]; dimers scFv [Gruber et al., J. Immunol. 152: 5368, 1994], linear antibodies [Zapata et al., Protein Eng. 8: 1057-62, 1995]; and chelating recombinant antibodies [Neri et al., J Mol Biol. 246: 367-73, 1995].

[0050] Proteins and non-proteinic agents can be conjugated to antibodies by methods known in the art. Coupling methods include direct binding, binding via covalently attached linkers and specific binding pairs (e.g. avidin-biotin). Such methods are, for example, those described by Greenfield et al., Cancer Research 50, 6600-6607 (1990) for the conjugation of doxorubicin and those described by Arnon et al., Adv. Exp. Med. Biol. 303, 79-90 (1991) and by Kiselev et al., Mol. Biol. (USSR) 25,508-514 (1991) for coupling platinum compounds.

[0051] In certain embodiments, antibodies and antibody fragments described herein are obtained, for example, from naturally occurring antibodies or ph to display phage Fab or scFv. The term "humanized antibody" refers to an antibody derived from a non-human antibody sequence, typically a monoclonal rodent antibody that contains modifications that make its sequence more similar to human. Alternatively, the humanized antibody may be derived from a chimeric antibody.

[0052] Antibody fragments comprise a domain antibody fragment (dAb) (Ward et al., Nature 341: 544-546, 1989) consisting of a VH domain, "linear antibodies" containing a pair of tandem segments Fd (VH-CH1-VH-CH1 ) forming a pair of antigen-binding regions. Linear antibodies

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EP 2 569 010 B1 may be bispecific or monospecific (Zapata et al., Protein Eng. 8: 1057-62 (1995)); A "minicide" consisting of scFv fused to CH3 via a peptide linker (no hinge) or via an IgG hinge is described in Olafsen et al., Protein Eng Des Sel. April 2004; 17 (4): 315-23; "Maximal" refers to divalent scFvs attached covalently to an Fc region of an immunoglobulin, see e.g.

Fredericks et al., Protein Engineering, Design & Selection, 17: 95-106 (2004) and Powers et al., Journal of Immunological Methods, 251: 123-135 (2001); heavy chain antibodies, e.g. VHH or H2L2 domain (referred to as "heavy chain" or "HCAb"); or camel VHH (see e.g.

Reichman et al., J. Immunol Methods 1999, 231: 25-38, Desmyter et al., J. Biol. Chem. 276: 26285-90, 2001, Ewert et al., Biochemistry 41: 3628-36, 2002, nanobodies (Cortez-Retamozo et al., Cancer Research 64: 2853-57, 2004); intracellular antibodies are single-chain antibodies expressed intracellularly and can manipulate the intracellular function of the protein (Biocca et al. EMBO J.

9: 101-108, 1990, Colby et al., Proc Natl Acad Sci USA 101: 17616-21, 2004, Mhashilkar et al., EMBO J 14: 1542-51, 1995, Wheeler et al. (FASEB J. 17 : 1733-5, 2003), trans-antibodies are antibodies that penetrate into cells in which the protein transduction domain (PTD) is conjugated to single chain variable fragment (scFv) antibodies to Heng et al. (Med Hypotheses. 64: 1105-8, 2005) , SMIP or immunoglobulin fusion proteins with a binding domain specific for the target protein are single-chain polypeptides containing antigen-binding domains linked to the immunoglobulin domains necessary to perform antibody effector functions. See, e.g., WO03 / 041600, U.S. Pat. Nos. 20030133939 and 20030118592.

II. Calcium salts and acetate salts or buffers [0053] It has been found that the addition of relatively low concentrations of calcium acetate to the preparations of the selected antibody reduces the viscosity of the formulation. The term "viscosity" as used herein refers to "absolute viscosity". The absolute viscosity, sometimes referred to as dynamic or straight viscosity, is the product of kinematic viscosity and liquid density: Absolute viscosity =₂ kinematic viscosity x density. The dimension of kinematic viscosity is L²/ T, where L is length and T is time. Usually, kinematic viscosity is expressed in centostats (cSt). SI unit of viscosity₂ kinematic is mm²/ si this is 1 cSt. The absolute viscosity is expressed in centipoise (cP).

The SI unit of the absolute viscosity is millipaskal times the second (mPa-s), where 1 cP = 1 mPa-s.

[0054] Such viscosity measurements can be made for hours (e.g., 1-23 hours), days (e.g., 1-10 days), weeks (e.g., 1-5 weeks) or months (e.g. 1-12 months) or years (e.g. e.g. 1-2 years, 1-3 years) after the addition of a viscosity reducing agent to the antibody preparation. The viscosity measurements can be carried out at the storage or feeding temperature, e.g. 2-8 ° C or 25 ° C (room temperature). In some embodiments, the absolute viscosity of the liquid or reconstituted liquid formulation at storage and / or administration temperature is 10, 9, 8, 7, 6, 5 or 4 cP or less.

[0055] In some embodiments, the viscosity of the protein preparation is measured before and after the addition of calcium acetate (and / or buffer). Viscosity measurement methods are well known in the art and include, for example, the use of a capillary viscometer or cone rheometer. Any method may be used, provided that the same method is used to compare the test and reference preparations.

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[0056] The viscosity of the antibody preparation can be reduced by adding a calcium salt and / or acetate salt (and / or buffer) to the formulation. The viscosity of the antibody preparation can be reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85% and about 90% compared to the viscosity of a comparable antibody preparation without a calcium salt and / or an acetate salt (and / or buffer).

[0057] Exemplary calcium salts include, but are not limited to, calcium acetate, calcium carbonate, and calcium chloride. In some embodiments, the calcium acetate has a concentration of at least 0.5 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM or 10 mM. In certain embodiments, the concentration of calcium salt is not greater than 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM or 20 mM. Any range formed by a combination of the above constraints, including, but not limited to, from about 0.5mM to about 10mM, about 5mM to about 10mM, or about 5mM to about 15mM is contemplated. In some embodiments, the calcium acetate is present at a concentration reducing the viscosity of the antibody preparation by at least 30%, 40%, 50%, 60% or more compared to the same acetate preparation and / or acetate buffer or which achieves a viscosity of 10 cP or less or 9, 8, 7, 6 or 5 cP or less. In certain embodiments, calcium acetate is added at low concentrations so as not to have a negative effect on the protein preparation. For example, at a concentration of calcium chloride or magnesium chloride, 20 mM or more of the protein can form a gel at low storage temperatures (e.g., 2-8 ° C). Consequently, in general, the concentration of calcium acetate at which the viscosity is reduced at the intended storage temperature of the reduced viscosity formulation is generally selected. not to have a negative effect on the protein preparation. For example, at a concentration of calcium chloride or magnesium chloride, 20 mM or more of the protein can form a gel at low storage temperatures (e.g., 2-8 ° C). Consequently, in general, the concentration of calcium acetate at which the viscosity is reduced at the intended storage temperature of the reduced viscosity formulation is generally selected. not to have a negative effect on the protein preparation. For example, at a concentration of calcium chloride or magnesium chloride, 20 mM or more of the protein can form a gel at low storage temperatures (e.g., 2-8 ° C). Consequently, in general, the concentration of calcium acetate at which the viscosity is reduced at the intended storage temperature of the reduced viscosity formulation is generally selected.

[0058] For all ranges described herein, the described concentration of a cation, anion or salt is the final concentration in the liquid or reconstituted liquid formulation to be administered. For any of the ranges described herein, range limitations are included in the range. However, the description also contemplates the same ranges for which the lower and / or upper limits are excluded from the range.

[0059] In some embodiments, the formulation described herein further comprises, in addition to calcium salt, an acetate buffer at a concentration of at least 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM or 15 MM. In some embodiments, the concentration is not greater than 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM or 50 mM. Any range determined by a combination of the above constraints, including but not limited to from about 5mM to about 15mM or from about 5mM to about 10mM, is contemplated. The buffer is preferably added at a concentration that maintains a pH of about 5-6 or 5-5.5 or 4.5-5.5. In some embodiments, the total acetate concentration is from about 10mM to about 50mM or from about 20mM to about 40mM.

[0060] In some aspects of the invention, the formulation comprises a total acetate concentration of at least about 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM or 50 mM. In some embodiments, the acetate concentration does not exceed 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM or 90 mM. Any range determined by a combination of the above constraints is contemplated, including but not limited to: from about 10mM to about 50mM, about 20mM to about 50mM, about 20mM to about 40mM, about 30mM to about 50mM or from about 30mM to about 75mM. In some embodiments, the acetate buffer is calcium acetate and / or sodium acetate. Alternatively, in certain embodiments, a salt and / or an acetate buffer

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EP 2 569 010 B1 is present at a concentration reducing the viscosity of the antibody preparation by at least 30%, 40%, 50%, 60% or more compared to the same antibody preparation with salt and / or acetate buffer or the formulation achieves a viscosity of 10 cP or less or 9, 8, 7, 6 or 5 cP or less. As a non-limiting example, a solution containing 10mM calcium acetate will contain 20mM acetate anion and 10mM calcium cation due to the divalence of the calcium cation, while the solution containing 10mM sodium acetate will contain 10mM sodium cation and 10mM acetate anion.

[0061] In some embodiments, the total concentration of ions (cations and anions) in the solution is at least 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55MM. , 60 mM, 65 mM, 70 mM, 75 mM, 80 mM or 85 mM. In some embodiments, the total ion concentration is not greater than about 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM , 95 mM, 100 mM, 110 mM, 120 mM, 130 mM, 140 mM, 150 mM, 160 mM, 170 mM, 180 mM, 190 mM or 200 mM. Any range constituted by a combination of the above constraints is contemplated, including but not limited to: from about 30mM to about 60mM or about 30mM to about 70mM, or about 30mM to about 80mM, or about 40mM to about 150mM or from about 50mM to about 150mM. As a non-limiting example,

[0062] For any of the formulations described herein, in some embodiments, the total osmolarity is not greater than 500 mOsm / L, 450 mOsm / L, 400 mOsm / L or 350 mOsm / L is preferably close to an isotonic osmolarity, e.g. 250-350 mOsm / l.

[0063] Other excipients known in the art or described herein may also be included in the formulation.

III. Excipients in the preparation [0064] In general, protein preparations are administered parenterally. When administered parenterally, they must be sterile. Sterile diluents include liquids that are pharmaceutically acceptable (safe and non-toxic when administered to humans) and suitable for the preparation of a liquid formulation such as a formulation reconstituted after lyophilization. Exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), pH buffered solution (e.g., phosphate buffered saline), sterile physiological saline, Ringer's solution, or dextrose solution. Diluents may include aqueous solutions of salts and / or buffers.

[0065] The excipients are additives introduced into the formulation because they either impart or improve the stability, delivery and ability to produce the medicinal product. Regardless of the reason for their inclusion, excipients are an integral part of the medicinal product and therefore must be safe and well tolerated by patients. In the case of protein drugs, the selection of excipients is particularly important because they can affect both the efficacy and immunogenicity of the drug. Therefore, protein preparations should be developed with the appropriate selection of excipients that will ensure adequate stability, safety and commercial value.

[0066] The excipients described herein are grouped according to their chemical type or their functional role in the formulations. When discussing each type of substance

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The short description of the stabilization methods is provided. Given the teachings and teachings provided herein, those skilled in the art will be able to easily vary the amount or range of excipient content without increasing viscosity to an undesirable level. Excipients may be selected to achieve the desired osmolarity (i.e., isotonicity, hypotonicity or hypertonicity) of the final solution, pH, desired stability, aggregation resistance or decomposition or precipitation, protection under freezing conditions, freeze drying or high temperatures or other properties. Various types of excipients are known. Exemplary excipients include salts, amino acids, other tonicity agents, surfactants, stabilizers, squeezing agents, cryoprotective agents,

[0067] Furthermore, if a particular excipient is administered in a formulation, e.g. in percent (wt)% w / v, it will be understood by those skilled in the art that an equivalent molar concentration of this excipient is also contemplated.

A. Buffers [0068] The pH range for optimum stability should be determined early in the preparation studies. A number of methods have proved useful in this undertaking, such as accelerated stability studies and calorimetric screening tests (Remmele RL Jr. et al., Biochemistry, 38 (16): 5241-7 (1999)). Once the composition of the preparation has been established, the medicinal product must be manufactured and maintained within the established storage limits throughout the storage period. Therefore, buffers are almost always used to control the pH in the formulation.

[0069] Organic acids, phosphates and tris were routinely used as buffers in the protein preparations (Table 1). Buffer capacity of buffering substances is maximum at pH equal to pKa and decreases with increasing or decreasing pH value from this value. In terms of one unit, around 90% of buffer capacity is maintained around pKa. Also, the buffer capacity increases in proportion to the increase in buffer concentration.

[0070] A number of factors have to be taken into account when selecting a buffer. First of all, the buffer substance and its concentration must be determined on the basis of its pKa and the desired pH of the formulation. It is equally important to ensure that the buffer is compatible with the protein drug, other excipients in the formulation and does not catalyze any degradation reactions. Recently, it has been demonstrated that polyanionic carboxylate buffers, such as citrate and succinate, form covalent adducts with protein side chain residues. The third important aspect to consider is the possible stinging and irritation that can cause the buffer. For example, citrate is known to cause a stinging sensation during an injection (Laursen T et al., Basic Clin Pharmacol Toxicol., 98 (2): 218-21 (2006)). The potential for inducing sting and irritation is greater for subcutaneous or intramuscular medications, where the drug solution stays at the site of administration for a longer period of time than when administered intravenously, where the preparation is rapidly diluted in the blood after administration. For preparations administered with a direct intravenous infusion, the total amount of buffer (and any other component of the preparation) should be monitored. For example, it has been reported that potassium ions administered in the form of a potassium phosphate buffer can For preparations administered with a direct intravenous infusion, the total amount of buffer (and any other component of the preparation) should be monitored. For example, it has been reported that potassium ions administered in the form of a potassium phosphate buffer can For preparations administered with a direct intravenous infusion, the total amount of buffer (and any other component of the preparation) should be monitored. For example, it has been reported that potassium ions administered in the form of a potassium phosphate buffer can

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This may result in cardiovascular reactions in the patient (Hollander-Rodriguez JC et al., Am. Fam Physician. 73 (2): 283-90 (2006)).

Table 1: Frequently used buffering agents and their pKa values

<td>Buffer</td><td>pKA</td><td>An example of a medicinal product</td>

<td>of acetate</td><td>4.8</td><td>Neupogen, Neulasta</td>

<td>succinate</td><td>pKa1 = 4.8, pKa2 = 5.5</td><td>Actimmune®</td>

<td>citrate</td><td>pKa1 = 3.1, pKa2 = 4.8, pKa3 = 6.4</td><td>Humira</td>

<td>Histidine (imidazole)</td><td>6.0</td><td>Xolair</td>

<td>phosphate</td><td>pKa1 = 2.15, pKa2 = 7.2, pKa3 = 12.3</td><td>Enbrel (liquid preparation)</td>

<td>tris</td><td>8.1</td><td>Leukine</td>

[0071] The buffer system present in the formulation is selected to be physiologically compatible and maintains the desired pH.

[0072] The pH buffering compound may be in any amount suitable to maintain the pH of the formulation at a predetermined level. The pH buffering agent, e.g. acetate, may be present at a concentration between 0.1 mM and 1000 mM (1 M). In one embodiment, the concentration of pH buffering agent is at least 0.1, 0.5, 0.7, 0.8, 0.9, 1.0, 1.2, 1.5, 1.7, 2, 3 , 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100 , 200, 500, 700 or 900 mM. In another embodiment, the pH of the pH buffering agent is between 1, 1.2, 1.5, 1.7, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80 or 90 mM and 100 mM. In yet another embodiment, the pH of the pH buffering agent is between 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30 or 40 mM and 50 mM .

[0073] Other exemplary pH buffering agents used to buffer the formulation presented herein include, but are not limited to, glycine, glutamate, succinate, phosphate, acetate and aspartate. Amino acids such as histidine and glutamic acid can also be used as buffering agents.

B. Stabilizers and Nailer substances [0074] Stabilizers are a class of compounds that can serve as cryoprotectants, freeze-thaw protection agents and glass forming agents. Cryoprotectants act as stabilizing proteins during freezing or in a frozen state at low temperatures. Lyophilization protection agents stabilize proteins in the lyophilized solid dosage form maintaining the native conformational properties of the protein during the dehydration steps during lyophilization. Properties of the vitreous state were classified as "strong" or "delicate" depending on the relaxation properties depending on the temperature. It is important that cryoprotectants, lyophilization protection agents and glass forming agents remain in the same phase with the protein in order to give durability. This category includes sugars,

[0075] The polyols include a group of excipients that includes sugars (e.g., mannitol, sucrose, sorbitol) and other polyhydric alcohols (e.g., glycerol and propylene glycol). This category includes

VP / 4572 / AGR

Polymer poly (ethylene glycol) (PEG). As stabilizing excipients and / or isotonizing agents, polyols are commonly used for parenteral protein preparations in both liquid and lyophilized form. Polyols can protect proteins against both physical and chemical degradation.

[0076] Exemplary C3-C6 polyols include propylene glycol, glycerol (glycerol), trerose, treitol, erythrosis, erythritol, ribose, arabinose, arabitol, lixx, maltitol, sorbitol, sorbose, glucose, mannose, mannitol, levulose, dextrose, maltose. , trehalose, fructose, xylitol, inositol, galactose, xylose, fructose, sucrose, 1,2,6-hexanetriol and the like. The sub-sugars include dextran, propylene glycol or polyethylene glycol. Reducing sugars, such as fructose, maltose or galactose, oxidize more readily than non-reducing sugars. Additional examples of sugar alcohols are glucitol, maltitol, lactitol or iso-maltulose. Additional exemplary protection agents for lyophilization include glycerol and gelatin, as well as sugars, mellibiosis, melezitosis, raffinose, mannotriosis and stachyose. Examples of reducing sugars include glucose, maltose, lactose, maltulose, iso-maltulose and lactulose. Examples of non-reducing sugar include non-reducing glycosides of polyhydroxy compounds selected from sugar alcohols and other straight chain polyols. Monoglycosides include compounds obtained by the reduction of disaccharides, such as lactose, maltose, lactulose and maltulose.

[0077] In certain embodiments, the formulations described herein also comprise a stabilizer (or a combination of stabilizers). The term "stabilizer" means an excipient capable of preventing aggregation or other physical degradation as well as chemical degradation (e.g. autolysis, deamidation, oxidation, etc.) in an aqueous and solid state. Stabilizers conventionally used in pharmaceutical compositions include, but are not limited to, sucrose, trehalose, mannose, maltose, lactose, glucose, raffinose, cellobiose, gentiobiose, isomaltose, arabinose, glucosamine, fructose, mannitol, sorbitol, glycine, arginine, HCL, polyhydroxyl compounds, including polysaccharides, such as dextran, starch, hydroxyethyl starch, cyclodextrins, N-methylpyrrolidene, cellulose and hyaluronic acid, sodium chloride [Carpenter et al. Develop. Biol. Standard 74: 225, (1991)]. In one embodiment, the stabilizer is incorporated at a concentration of about 0% to about 40% w / v. In another embodiment, the stabilizer is introduced at a concentration of at least 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30 or 40% w / v. In another embodiment, the stabilizer is incorporated at a concentration of about 1, 2, 3, 4, 5, 6, 7, 8, 9% to about 10% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 2% to about 6% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 4% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 6% w / v. In one embodiment, the stabilizer is incorporated at a concentration of about 0% to about 40% w / v. In another embodiment, the stabilizer is introduced at a concentration of at least 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30 or 40% w / v. In another embodiment, the stabilizer is incorporated at a concentration of about 1, 2, 3, 4, 5, 6, 7, 8, 9% to about 10% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 2% to about 6% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 4% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 6% w / v. In one embodiment, the stabilizer is incorporated at a concentration of about 0% to about 40% w / v. In another embodiment, the stabilizer is introduced at a concentration of at least 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30 or 40% w / v. In another embodiment, the stabilizer is incorporated at a concentration of about 1, 2, 3, 4, 5, 6, 7, 8, 9% to about 10% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 2% to about 6% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 4% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 6% w / v. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30 or 40% w / v. In another embodiment, the stabilizer is incorporated at a concentration of about 1, 2, 3, 4, 5, 6, 7, 8, 9% to about 10% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 2% to about 6% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 4% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 6% w / v. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30 or 40% w / v. In another embodiment, the stabilizer is incorporated at a concentration of about 1, 2, 3, 4, 5, 6, 7, 8, 9% to about 10% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 2% to about 6% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 4% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 6% w / v. the stabilizer is introduced at a concentration of about 4% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 6% w / v. the stabilizer is introduced at a concentration of about 4% w / v. In yet another embodiment, the stabilizer is incorporated at a concentration of about 6% w / v.

[0078] If desired, the formulations also contain suitable amounts of nailing agents and osmolarity regulators suitable for forming a lyophilized "cake". Nucleating agents can be crystalline (e.g. mannitol, glycine) or amorphous (e.g. sucrose, polymers such as dextran, polyvinylpyrrolidone, carboxymethylcellulose). Other exemplary naive agents include lactose, sorbitol, trehalose and xylitol. In a further embodiment, the tamping agent is introduced at a concentration of about 0% to about 10% w / v. In another embodiment, the tamping agent is introduced at a concentration of at least 0.2, 0.5, 0.7, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4, 0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0 or 9.5% w / v . In yet another embodiment, the nailing agent is

VP / 4572 / AGR

It is present in a concentration of about 1, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5% to 5.0% w / v. for the preparation of a mechanically and pharmaceutically stable cake.

C. Surfactants [0079] Protein molecules have a high tendency to interact with surfaces, making them susceptible to adsorption and denaturation at the interface between air and liquid, liquid and the vial and liquid and liquid (silicone oil). It has been observed that this degradation pathway is inversely dependent on protein concentration and results in the formation of soluble and insoluble protein aggregates or the loss of protein from solution by adsorption on surfaces. In addition to adsorption on the surface of the vessel, the surface-induced degradation is exacerbated by physical shaking, as is the case when transporting and handling the product.

[0080] Surfactants are commonly used in protein preparations to prevent surface-induced degradation. Surfactants are amphipathic molecules with the ability to compete with proteins for a phase boundary position. The hydrophobic portions of surfactant molecules occupy interfacial positions (e.g., air / liquid), while the hydrophilic portions of the molecules remain oriented towards the depth of the solvent. At sufficient concentrations (usually near the critical concentration of the micellar detergent), the surface layer of surfactant molecules prevents the adsorption of protein particles on the surface. In this way, surface-induced degradation is minimized. The most commonly used surfactants are esters of fatty acids and sorbitan polyethoxylates, i.e. polysorbate 20 and polysorbate 80 (e.g., Avonex®, Neupogen®, Neulasta®). These two differ only in the length of the aliphatic chain, C-12 and C-18, respectively, which gives the molecules hydrophobic properties. Accordingly, polysorbate 80 is more surface-active and has lower critical micelle concentration than polysorbate 20. A number of commercial liquid products such as Gonal-F®, Norditropin® and Ovidrel® also used poloxamer 188 surfactant.

[0081] Detergents can also affect the thermodynamic conformational stability of proteins. Again, the effects of a given excipient will be specific for the protein. For example, polysorbates have been shown to reduce the stability of certain proteins and increase the stability of others. The destabilization of detergent proteins can be explained by the specific interaction of the hydrophobic tails of detergent molecules with partially or fully developed forms of the protein. These types of interactions could cause a shift in the conformational equilibrium towards more spatially expanded protein states (i.e., increasing the exposure of the hydrophobic portions of the protein molecule due to complementarity with the binding polysorbate). Alternatively, if the native form of the protein has certain hydrophobic surfaces,

[0082] Another aspect of the polysorbates is that they are inherently susceptible to oxidative degradation. Often, as raw materials, they contain sufficient amounts of peroxides to cause oxidation of the side chains of protein residues, especially methionine. The ability to oxidative damage after the addition of a stabilizer emphasizes that the lowest effective concentrations of excipients should be used in the formulations. In the case of surfactants, the effective concentration for a given protein depends on the mechanism of stabilization. It was postulated that if the mechanism stabilizes the measure

VP / 4572 / AGR

As the surfactant is associated with the prevention of surface denaturation, the effective concentration will fall close to the critical concentration of the micellar detergent. In contrast, if the stabilization mechanism is associated with specific protein-detergent interactions, the effective concentration of surfactant will depend on protein concentration and the stoichiometry of interaction (Randolph TW et al., Pharm Biotechnol., 13: 159-75 (2002)).

[0083] Surfactants can also be added in amounts suitable to prevent the phenomenon of surface aggregation during freezing and drying [Chang, B, J. Pharm. Sci. 85: 1325, (1996)]. Exemplary surfactants include anionic, cationic, nonionic, zwitterionic and amphoteric surfactants, including surfactants derived from naturally occurring amino acids. Anionic surfactants include, but are not limited to, sodium lauryl sulfate, sodium dioctylsulfosuccinate and sodium dioctylsulfonate, chenodeoxycholic acid, sodium N-lauroyl sarcosine, lithium dodecyl sulfate, 1-octanesulfonic acid sodium salt, sodium hydrate hydrate, sodium deoxycholate, and glycodezoxycholic acid sodium salt. Cationic surfactants include, but are not limited to, benzalkonium chloride or benzethonium chloride, monohydrous cetylpyridinium chloride and hexadecyltrimethylammonium bromide. The zwitterionic surfactants include, but are not limited to, CHAPS, CHAPSO, SB3-10, and SB3-12. Non-ionic surfactants include, but are not limited to, digitonin, Triton X-100, Triton X-114, TWEEN-20 and TWEEN-80. In another embodiment, the surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene, hydrogenated castor oil 10, 40, 50 and 60, glyceryl monostearate, polysorbate 40, 60, 65 and 80, soybean lecithin and other phospholipids, such as DOPC, DMPG, DMPC and DOPG; fatty acid ester of sucrose, methylcellulose and carboxymethylcellulose. The zwitterionic surfactants include, but are not limited to, CHAPS, CHAPSO, SB3-10, and SB3-12. Non-ionic surfactants include, but are not limited to, digitonin, Triton X-100, Triton X-114, TWEEN-20 and TWEEN-80. In another embodiment, the surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene, hydrogenated castor oil 10, 40, 50 and 60, glyceryl monostearate, polysorbate 40, 60, 65 and 80, soybean lecithin and other phospholipids, such as DOPC, DMPG, DMPC and DOPG; fatty acid ester of sucrose, methylcellulose and carboxymethylcellulose. The zwitterionic surfactants include, but are not limited to, CHAPS, CHAPSO, SB3-10, and SB3-12. Non-ionic surfactants include, but are not limited to, digitonin, Triton X-100, Triton X-114, TWEEN-20 and TWEEN-80. In another embodiment, the surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene, hydrogenated castor oil 10, 40, 50 and 60, glyceryl monostearate, polysorbate 40, 60, 65 and 80, soybean lecithin and other phospholipids, such as DOPC, DMPG, DMPC and DOPG; fatty acid ester of sucrose, methylcellulose and carboxymethylcellulose. In another embodiment, the surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene, hydrogenated castor oil 10, 40, 50 and 60, glyceryl monostearate, polysorbate 40, 60, 65 and 80, soybean lecithin and other phospholipids, such as DOPC, DMPG, DMPC and DOPG; fatty acid ester of sucrose, methylcellulose and carboxymethylcellulose. In another embodiment, the surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene, hydrogenated castor oil 10, 40, 50 and 60, glyceryl monostearate, polysorbate 40, 60, 65 and 80, soybean lecithin and other phospholipids, such as DOPC, DMPG, DMPC and DOPG; fatty acid ester of sucrose, methylcellulose and carboxymethylcellulose.

[0084] The formulations described herein may further comprise these surfactants alone or as a mixture in varying proportions. In one embodiment, the surfactant is incorporated at a concentration of about 0% to about 5% w / v. In another embodiment, the surfactant is incorporated in a concentration of at least 0.001, 0.002, 0.005, 0.007, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0 or 4, 5% w / v In another embodiment, the surfactant is incorporated at a concentration of about 0.001% to about 0.5% w / v. In yet another embodiment, the surfactant is incorporated at a concentration of about 0.004, 0.005, 0.007, 0.01, 0.05 or 0.1% w / v. up to about 0.2% w / v. In yet another embodiment,

[0085] In some embodiments, the viscosity reduction is achieved with a relatively small addition of or without the surfactant, e.g. 0.1% or less of total surfactant, or 0.05% or less or 0.01% or less.

D. Amino acids [0086] Amino acids have found universal use in protein preparations as buffers, tussors, stabilizers and antioxidants. For the buffering of protein preparations in the pH range 5.5-6.5 and 4.0-5.5, histidine and glutamic acid, respectively, are used. The imidazole group of histidine has pKa =

6.0, and the carboxyl group of the glutamic acid side chain has a pKa of 4.3, which makes them

VP / 4572 / AGR

EP 2 569 010 B1 suitable for buffering at appropriate pH ranges. Glutamic acid is used in some preparations (e.g., Stemgen®). Histidine is commonly present in commercially available protein preparations (e.g., Xolair®, Herceptin®, Recombinate®). It is a good alternative to citrate, a buffer known to cause stinging sensation after injection. Interestingly, histidine also has a stabilizing effect when used in high concentrations in both liquid and lyophilized forms (Chen B et al., Pharm Res., 20 (12): 1952-60 (2003)). It was also observed that histidine (up to 60 mM) reduces the viscosity of the preparation with high concentration of this antibody. However, in the same study, the authors observed increased aggregation and discoloration in preparations containing histidine during freezing and thawing tests of the antibody in stainless steel containers. The authors attributed this to the effect of iron ions eluted due to the corrosion of steel containers. Another caution regarding the use of histidine is that it oxidizes in the presence of metal ions (Tomita M. et al., Biochemistry, 8 (12): 5149-60 (1969)). It seems promising to use methionine as an antioxidant in preparations; it has been observed that it is effective in the prevention of a number of oxidative hazards (Lam XM et al., J Pharm Sci., 86 (11): 1250-5 (1997)). as for caution when using histidine, it is oxidized in the presence of metal ions (Tomita M. et al., Biochemistry, 8 (12): 5149-60 (1969)). It seems promising to use methionine as an antioxidant in preparations; it has been observed that it is effective in the prevention of a number of oxidative hazards (Lam XM et al., J Pharm Sci., 86 (11): 1250-5 (1997)). as for caution when using histidine, it is oxidized in the presence of metal ions (Tomita M. et al., Biochemistry, 8 (12): 5149-60 (1969)). It seems promising to use methionine as an antioxidant in preparations; it has been observed that it is effective in the prevention of a number of oxidative hazards (Lam XM et al., J Pharm Sci., 86 (11): 1250-5 (1997)).

[0087] Amino acids glycine, proline, serine and alanine stabilize proteins. Glycine is also a commonly used squeezing agent in lyophilized formulations (e.g., Neumega®, Genotropin®, Humatrope®). Arginine has been shown to be an effective aggregation inhibiting agent and has been used in both liquid and lyophilized preparations (e.g. Activase®, Avonex®, Enbrel® liquid).

E. Antioxidants [0088] Oxidation of protein residues takes place for a variety of reasons. In addition to the addition of specific antioxidants, the prevention of oxidative protein damage includes careful control of a number of factors during the product manufacturing and storage process, such as oxygen in the atmosphere, temperature, exposure to light and chemical contamination. The most commonly used pharmaceutical antioxidants are reducing agents, oxygen scavengers / free radicals, or chelating agents. Antioxidants in therapeutic protein preparations must be water-soluble and remain active throughout the shelf life of the product. Oxygen reducing agents and scavengers / free radicals act by removing the active oxygen species from the solution. Chelating agents, such as EDTA, they can be effective thanks to the binding of trace metallic impurities conducive to the formation of free radicals. For example, EDTA was used in a liquid formulation of acid fibroblast growth factor to inhibit oxidation of metal-catalyzed cysteine ​​residues. EDTA has been used in commercial products such as Kineret® and Ontak®.

[0089] However, antioxidants can themselves induce other covalent or physical changes in the protein.

Many such cases have been described in the literature. Reducing agents (such as glutathione) can cause the breakdown of intramolecular disulfide bonds, which may result in disulfide shuffling. It has been shown that in the presence of transition metal ions, ascorbic acid and EDTA promote the oxidation of methionine in a number of proteins and peptides (Akers MJ and Defelippis MR Peptides and Proteins as Parenteral Solutions in Pharmaceutical Formulation Development of Peptides and Proteins.) Sven Frokjaer, Lars Hovgaard, ed. : Pharmaceutical Science, Taylor and Francis, United Kingdom (1999)); Fransson JR, J. Pharm. Sci. 86 (9): 4046-1050 (1997); Yin J. et al., Pharm Res., 21 (12): 2377-83 (2004)). It was reported that

VP / 4572 / AGR

Sodium thiosulphate reduces the level of light and temperature induced oxidation of methionine in the case of rhuMab HER2; however, this study also reported the formation of a thiosulphate-protein adduct (Lam XM, Yang JY et al., J Pharm Sci. 86 (11): 1250-5 (1997)). A suitable antioxidant is selected taking into account the specific hazards and the sensitivity of the protein.

F. Metal ions [0090] In general, transition metal ions are undesirable in protein preparations because they can catalyze physical and chemical degradation reactions of proteins. However, certain metal ions are included in the formulations when they are cofactors of proteins and for suspension preparations of proteins in which they form coordination complexes (e.g., a zinc suspension of insulin). Recently, the use of magnesium ions (10-120 mM) has been proposed to inhibit the isomerization of aspartic acid to isoaspartamic acid (WO 2004/039337).

[0091] Two examples in which metal ions provide stability or increased activity of proteins are human deoxyribonuclease (rhDNase, Pulmozyme®) and factor VIII. In the case of rhDNase, Ca ions²⁺(up to 100 mM) increased the stability of the enzyme via a specific binding site (Chen B. et al., J Pharm Sci., 88 (4): 477-82 (1999)). Indeed, the removal of calcium ions from the solution by EGTA resulted in increased deamidation and aggregation. However, this effect was observed only in the case of Ca ions²⁺; it was observed that other divalent cations - Mg²⁺, Mn²⁺ and Zn²⁺ they cause the destabilization of rhDNase. Similar effects were observed for factor VIII. Ions Ca²⁺ and Sr.²⁺ they stabilized the protein while others, such as Mg²⁺, Mn²⁺ and Zn²⁺, Cu²⁺ and Fe²⁺ they caused destabilization of the enzyme (Fatouros, A. et al., Int. J. Pharm., 155, 121-131 (1997)). In a separate study with factor VIII, a significant increase in the rate of aggregation in the presence of Al ions was observed³⁺ (Derrick, TS et al., J. Pharm. Sci., 93 (10): 2549-57 (2004)). The authors note that other excipients, such as buffer salts, are often contaminated with Al ions³⁺, which indicates the need to use excipients of appropriate quality in preparations.

G. Preservatives [0092] Preservatives are necessary in the development of parenteral formulations intended for multiple use, which require more than one withdrawal from the same container. Their primary function is to inhibit the growth of microorganisms and ensure the sterility of the product throughout the storage period or period of use of the medicinal product. Common preservatives include phenol, benzyl alcohol, metacresol, alkyl parabens, such as methyl paraben or propyl paraben, benzalkonium chloride and benzethonium chloride. Other examples of compounds with antimicrobial activity of preservatives include octadecyldimethylbenzylammonium chloride, hexamethonium chloride. Other types of preservatives include aromatic alcohols, such as butyl alcohol, phenol, benzyl alcohol; atechol, resorcinol, cyclohexanol, 3-pentanol. Although preservatives have a long history of use, the development of protein preparations containing preservatives can be a challenge. Preservatives almost always have a destabilizing (aggregating) effect on proteins and this has become a major limiting factor in their use in preparations containing multiple doses of protein (Roy S. et al., J Pharm Sci., 94 (2): 38296 (2005)).

VP / 4572 / AGR

[0093] Drug presentations in the form of reusable pens contain preservative formulations. For example, currently available hGH preparations are available on the market. Norditropin® (liquid, Novo

Nordisk), Nutropin AQ® (liquid, Genentech) and Genotropin (freeze-dried, two-chamber container, Pharmacia &amp; Upjohn) contain phenol, while Somatrope® (Eli Lilly) was prepared using mkrezol.

[0094] During the development of the formulation in the form of a preserved dosage form, a number of aspects should be considered. The concentration of the preservative in the medicinal product should be optimized to make it effective. This requires testing a given preservative in the dosage form for concentration ranges that impart antibacterial efficacy without impairing the stability of the protein. For example, in the development of a liquid interleukin-1 receptor (type I) formulation, three preservatives were successfully tested using differential scanning calorimetry (DSC). Preservatives were ranked according to their effect on stability at concentrations commonly used in commercial products (Remmele RL Jr. et al. Pharm Res., 15 (2): 200-8 (1998)).

[0095] Some preservatives can cause reactions at the injection site, which is another factor to consider when choosing a preservative. In clinical trials that focused on the evaluation of preservatives and buffers in Norditropin, pain was found to be lower for formulations containing phenol and benzyl alcohol compared to the m-cresol formulation (Kappelgaard AM, Horm Res. 62 added 3 : 98-103 (2004)). Interestingly, among the commonly used preservatives, benzyl alcohol has anesthetic properties (Minogue SC and Sun DA, Anesth Analg., 100 (3): 683-6 (2005)).

IV. Kits [0096] As an additional aspect of the present disclosure, sets are described herein comprising one or more preparations described herein packaged in a manner that facilitates their use for administration to patients. Such a kit may contain a formulation as described herein (e.g., a composition comprising any of the antibodies described herein) packaged in a container, such as a closed bottle, vessel, single use or reusable vial, pre-filled syringe or pre-filled injection device, optionally with a label attached to the container or included in the package describing the use of the compound or composition in the practice of the method. In one aspect of this disclosure, the compound or composition is packaged in a unit dosage form. The kit may further comprise a device suitable for administering the composition according to a particular route of administration. Preferably, the kit includes a label that describes the use of the antibody described herein or the formulation described herein.

V. Dosage [0097] The dosage regimen of the method for treating the condition described herein should be determined by the attending physician, taking into account various factors that modify the effect of the drugs, e.g., age, condition, weight, sex and diet of the patient, severity of possible infections , time of administration and other clinical factors. In various aspects, the daily dose is in the range of 0.1-50 mg of the antibody preparation per kilogram of body weight (taking into account the mass of the protein itself, without modification

VP / 4572 / AGR

EP 2 569 010 B1). In some embodiments, the dosage is about 0.5 mg / kg to 20 mg / kg or about 0.5-10 mg / kg.

[0098] Said formulations are generally administered parenterally, e.g. intravenously, subcutaneously, intramuscularly, via an aerosol (pulmonary or inhalation) or using a depot for sustained release. In some embodiments, the formulation is administered intravenously using an initial bolus followed by continuous infusion to maintain therapeutic levels of the medicinal product in the circulation. In other embodiments, the formulation is administered in a single dose. Normal specialists in the field will easily optimize effective dosages and delivery schedules that result from good medical practice and the clinical status of the patient. The frequency of dosing depends on the pharmacokinetic parameters of the agents and route of administration. The optimal pharmaceutical formulation will be determined by a person skilled in the art depending on the route of administration and the desired dose. See, for example, Remington's Pharmaceutical Sciences, ed. 18th (1990, Mack Publishing Co., Easton, Pennsylvania 18042) pp. 1435-1712. The form of such formulations may affect the physical state, stability, in vivo release rate and in vivo clearance rate of the agents administered. Depending on the route of administration, the appropriate dose can be calculated depending on the body weight, body surface or size of the organ. Further refinement of the calculations necessary to determine the appropriate treatment dose for each of the above-mentioned preparations is routinely performed by those skilled in the art without undue experimentation, especially in light of the dosage and testing information disclosed herein, as well as pharmacokinetic data obtained in the human clinical studies discussed above. Appropriate dosages can be determined using established blood dose assays in combination with appropriate dose-response data. The final dosing regimen will be determined by the attending physician taking into account various factors that modify the effect of the drugs, e.g. drug specific activity, severity of damage and patient response, age, condition, weight, sex and diet of the patient, severity of possible infections, time of administration and other clinical factors. As the research progresses, there will be additional information on the appropriate dosage levels and treatment time of various diseases and conditions.

VI. Therapeutic use of the preparation [0099] The formulations described herein are useful for treating or preventing bone related disorders, such as bone disorders associated with abnormal osteoblast or osteoclast activity. In some embodiments, the formulation is administered to a subject suffering from a bone disorder selected from the group consisting of achondroplasia, clavicular cystadenostasis, chondrichoma, fibrous dysplasia, Gaucher disease, hypophosphatemic rickets, Marfan syndrome, multiple inherited exostoses, neurofibromatosis, congenital bone fragility , osteopetrosis, osteopoikilii, atherosclerotic lesions, osteoarthritis, pyogenic osteoarthritis, periodontal disease,

VP / 4572 / AGR

Meloreostosis, metabolic bone diseases, mastocytosis, anemia / sickle cell disease, bone loss associated with organ transplants, bone loss associated with kidney transplantation, systemic lupus erythematosus, ankylosing spondylitis, epilepsy, ankylosing spondylitis , epilepsy, juvenile arthritis, thalassemia, mucopolysaccharidosis, Fabry's disease, Turner's syndrome, Down's syndrome, Klinefelter's syndrome, leprosy, Perthe's disease, idiopathic scoliosis of adolescents, which starts in childhood as a multi-system inflammatory disease, Winchester syndrome, Menkes' disease, Wilson's disease, ischemic bone disease (e.g., Legg-Calve-Perthes disease or local migrating osteoporosis), anemia,steroids induced, glucocorticoid-induced bone loss, heparin-induced bone loss, bone marrow disorders, scurvy, malnutrition, calcium deficiency, osteoporosis, osteopenia, alcoholism, chronic liver disease, postmenopausal state, chronic inflammation, rheumatoid arthritis, inflammatory disease intestine, ulcerative colitis, colitis, Crohn's disease, menstrual disorders, amenorrhea, pregnancy, diabetes, hyperthyroidism, thyroid disorders, parathyroid disorder, Cushing's disease, acromegaly, hypogonadism, immobilization or non-use, sympathetic dystrophy, localized osteoporosis, osteomalacia, bone loss associated with joint replacement, bone loss associated with HIV,bone loss associated with loss of growth hormone, loss of bone tissue associated with cystic fibrosis, bone loss associated with chemotherapy, tumor-induced bone loss, cancer-related bone loss, hormonal ablative bone loss, multiple myeloma, bone loss induced by drugs, anorexia nervosa, disease associated with bone loss of the face, associated with the disease of cranial bone loss, associated with bone osteonecrosis disease, cranial bone loss disease, loss of bone associated with aging, age-related facial loss, associated with age of cranial bone loss, age-related bone loss in the jaw, age-related cranial bone loss or loss of bone associated with weightlessness.

[0100] In some embodiments, the formulations described herein are useful for improving results in orthopedic procedures, dental procedures, implant surgery, articular replacement, bone transplantation, bone plastic surgery and bone repair, such as healing cracks, healing of non-growing parts , healing of delayed union and facial reconstruction. One or more compositions may be administered prior to, during and / or after surgery, replacement, transplantation, surgery or repair.

[0101] In order to obtain a beneficial biological response, such a preparation does not have to cure an individual from the disorder or completely protect him from the occurrence of a bone disorder. The preparation can be used prophylactically, which means full or partial protection against a bone related disorder or its symptom. The preparation may also be used therapeutically to fully or partially improve the state of the disorder or symptoms associated with the bones or to provide total or partial protection against further progression of the bone disorder or its symptoms.

Indeed, the materials and methods of the invention are particularly useful in increasing bone mineral density and maintaining increased bone mineral density over a period of time.

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[0102] One or more administrations of the formulation described herein can be performed in a therapeutic period that last for example from about 1 month to about 12 months (e.g., about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months or about 11 months). In some embodiments, the patient is administered one or more doses of the formulation to maintain bone mineral density. The term "maintaining bone mineral density" as used herein means that the increased bone mineral density obtained from the initial dose of the formulation does not fall by more than about 1% to about 5% in about 6 months, about 9 months, about 1 year, about 18 months, about 2 years or throughout the life of the patient. It should be noted that the patient may require alternating treatment phases to increase bone density and maintain bone density.

[0103] Furthermore, it may be desirable to administer multiple doses of the preparation or to divide the dosages over time depending upon the therapeutic regimen selected for the particular patient. The preparation can be administered periodically for one year or less (eg 9 months or less, 6 months or less, or 3 months or less). Accordingly, the formulation may be administered to a human once every 7 days or 2 weeks or 3 weeks or 1 month or 5 weeks or 6 weeks or 7 weeks or 2 months or 9 weeks or 10 weeks or 11 weeks or 3 months, 13 weeks or 14 weeks or 15 weeks or 4 months or 17 weeks or 18 weeks or 19 weeks or 5 months or 21 weeks or 22 weeks or 23 weeks or 6 months or 12 months.

VII. Combination Therapy [0104] Treating pathology by combining two or more agents targeting the same pathogen or biochemical pathway sometimes results in greater efficacy and reduced side effects relative to the use of a therapeutically effective dose of each of these drugs. In some cases the effectiveness of the drug combination is additive (the effectiveness of the combination is approximately equal to the sum of the effects of each drug administered alone), but in other cases the effect may be synergistic (the effectiveness of the combination is greater than the sum of the effects of each drug administered alone). The term "combination therapy" as used herein means that two compounds may be delivered in a simultaneous manner, e.g. simultaneously or in such a way that one of the compounds is administered first, and then the second agent is administered, e.g. sequentially. The desired result may be a subjective relief in one or more symptoms or an identifiable improvement in the recipient of the dose.

[0105] In some embodiments, the formulation is administered with a standard of therapeutic care to treat reduced bone mineral density. The term "standard care" as used herein refers to a treatment that is commonly accepted by clinicians for a particular type of patient who has been diagnosed with a particular type of disease. In some embodiments, the standard care drug is selected from the group consisting of an antiresorptive agent, an osteogenic agent, an estrogen receptor antagonist (including, but not limited to, raloxifene, bazedoxifene and lasofoxifene) and a drug that has a stimulatory effect on osteoclasts. In some embodiments, the anti-resorptive drug includes, but is not limited to, a bisphosphonate (including but not limited to alendronate, risedronate, ibandronate, and zoledronate),

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EP 2 569 010 B1 is a hormone replacement therapy. In some embodiments, the osteogenic agent comprises, but is not limited to, parathyroid hormone (PTH) or its peptide fragment, PTH-associated protein (PTHrp), bone morphogenetic protein, osteogenin, NaF, PGE2 agonist, statin and RANK ligand (RANKL). In some embodiments, the osteoclast-stimulant drug comprises, but is not limited to, vitamin D or a vitamin D derivative or substance that mimics it.

[0106] In certain embodiments, the formulation is administered to a patient in case the treatment with the standard care drug described herein is contraindicated.

EXAMPLES

Example 1 - Calcium acetate reduced the effective viscosity of sclerostin antibody preparations [0107] 10 ml of the selected anti-sclerostin antibody (75.7 mg / ml) were dialyzed against 2 liters of 10 mM Na (OAc) and 9% sucrose at 4 ° C for 2 hours. . The selected anti-sclerostin antibody (75.7 mg / ml) was concentrated to about 160 mg / ml and diluted with water to approximately 140 mg / ml and 120 mg / ml. The absorbance of the diluted samples was 120, 142 and 157 mg / ml, respectively.

[0108] 10 μl of 1.0 M Ca (OAc)₂ added to 1 ml of 120 mg / ml, 140 mg / ml and 160 mg / ml samples. Absolute viscosity, pH and osmolarity of samples were determined (see Table 2). The absolute viscosity of the samples (500 μΓ) was measured using a Brookfield LV-DVII cone-plate viscometer with a CPE-40 spindle with a temperature-adjusted sample cup controlled by circulation from a water bath with a constant temperature of 25 ° C.

Table 2

<td>A sample</td><td>Viscosity (cP)</td><td>pH</td><td>osmolarity</td>

<td>120 mg / ml (control)</td><td>18</td><td>5.3</td><td>375</td>

<td>120 mg / ml + 10 mM Ca (OAc)₂</td><td>8.4</td><td>5.4</td><td>398</td>

<td>142 mg / ml + 10 mM Ca (OAc)₂</td><td>17</td><td>5.4</td><td>450</td>

<td>157 mg / ml + 10 mM Ca (OAc)₂</td><td>36</td><td>5.4</td><td>610</td>

[0109] The results indicate that 10 mM Ca (OAc) 2 injected into the liquid composition of the selected antibody reduced the viscosity by about half. This experiment was carried out for each of Ab-4, Ab-5, Ab-13, Ab-14, Ab-19, Ab-20 and Ab-23 antibodies.

Example 2 - Adaptation of preparations [0110] 10 ml of the selected anti-sclerostin antibody (75.7 mg / ml) were dialyzed against 2 liters of 10 mM Na (OAc), 6% sucrose or 4% sucrose at 4 ° C for 2 hours. Each of the sucrose preparations was then concentrated using Amicons to about 140 mg / ml, and then diluted with water back to the target concentrations (i.e., 120 mg / ml, 140 mg / ml and 160 mg / ml). The absorbance of the diluted samples was 124 mg / ml (4% sucrose), 119.5 mg / ml (6% sucrose), 137.5 mg / ml (4% sucrose) and 142 mg / ml (6% sucrose, respectively). ).

[0111] 10 ml of 1.0 M Ca (OAc) was added to 1 ml of samples.₂. The viscosity, osmolality and pH of the samples were assessed (see Table 3)

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Table 3

<td>A sample</td><td>mM</td><td>mg / ml</td><td>pH</td><td>osmolarity</td><td>Viscosity (cP)</td>

<td>120 mg / ml + 10 mM CaOAC + 4% sucrose</td><td>10</td><td>124</td><td>5,285</td><td>214</td><td>6.2</td>

<td>120 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>119.5</td><td>5.25</td><td>282</td><td>5.7</td>

<td>140 mg / ml + 10 mM CaOAC + 4% sucrose</td><td>10</td><td>137.5</td><td>5,303</td><td>231</td><td>9.5</td>

<td>140 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>142</td><td>5.307</td><td>294</td><td>11</td>

[0112] The test was repeated as follows: 10 ml of the selected anti-sclerostin antibody (75.7 mg / ml) were dialyzed against 2 liters of 10 mM Na (OAc), 6% sucrose or 4% sucrose at 4 ° C for 2 hours. Each of the sucrose preparations was then concentrated using an Amicon filter to about 140 mg / ml, and then diluted with water back to the target concentrations (i.e., 70 mg / ml, 100 mg / ml and 120 mg / ml). The absorbance of the diluted samples was 71 mg / ml (4% sucrose), 68.2 mg / ml (6% sucrose), 99.4 mg / ml (4% sucrose), 100.5 (6% sucrose), respectively. , 122 mg / ml (4% sucrose) and 113 mg / ml (6% sucrose).

[0113] The pH, osmolality and viscosity of the samples were determined. See table 4.

Table 4

<td>A sample</td><td>mM</td><td>mg / ml</td><td>pH</td><td>osmolarity</td><td>Viscosity (cP)</td>

<td>70 mg / ml + 4% sucrose</td><td>10</td><td>71</td><td>5.205</td><td>154</td><td>3.5</td>

<td>70 mg / ml + 10 mM CaOAC + 4% sucrose</td><td>10</td><td>71</td><td>5.233</td><td>183</td><td>2.2</td>

<td>70 mg / ml 6% sucrose</td><td>10</td><td>68.2</td><td>5.201</td><td>231</td><td>3.4</td>

<td>70 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>68.2</td><td>5,279</td><td>256</td><td>2.4</td>

<td>100 mg / ml + 4% sucrose</td><td>10</td><td>99.4</td><td>5,265</td><td>165</td><td>8.1</td>

<td>100 mg / ml + 10 mM CaOAC + 4% sucrose</td><td>10</td><td>99.4</td><td>5.288</td><td>191</td><td>4.1</td>

<td>100 mg / ml + 6% sucrose</td><td>10</td><td>100.5</td><td>5.273</td><td>241</td><td>8.4</td>

<td>100 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>100.5</td><td>5,303</td><td>270</td><td>4.3</td>

<td>120 mg / ml + 4% sucrose</td><td>10</td><td>122</td><td>5,295</td><td>177</td><td>15.6</td>

<td>120 mg / ml + 10 mM CaOAC + 4% sucrose</td><td>10</td><td>122</td><td>5.306</td><td>202</td><td>6.9</td>

<td>120 mg / ml + 6% sucrose</td><td>10</td><td>113</td><td>5.3</td><td>249</td><td>15.4</td>

<td>120 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>113</td><td>5.311</td><td>274</td><td>6.6</td>

[0114] Lowering the pH of the Ca (OAc) 2 to 5.2 buffer allowed the pH of all final formulations to be maintained between 5.25 and 5.307. Formulations with 4% sucrose were below the isotonic range (250-350 mOsm / kg), but the preparations with 6% sucrose were in the middle of the isotonic range.

[0115] To further evaluate the effect of 6% sucrose and 10 mM Ca (OAc) 2 on viscosity reduction, the above test was repeated with consecutive concentrations of sclerostin antibody up to 160 mg / ml.

[0116] Samples were prepared as described above in the following concentrations: 120 mg / ml, 140 mg / ml and 160 mg / ml. 10 μΐ 1.0 M Ca (OAc) was added to each of the samples₂ at pH 5.2. The pH, osmolarity and viscosity of the samples were determined. See table 5.

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Table 5

<td>A sample</td><td>mM</td><td>mg / ml</td><td>pH</td><td>osmolarity</td><td>Viscosity (cP)</td>

<td>100 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>107</td><td>5,285</td><td>271</td><td>4.3</td>

<td>100 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>107</td><td>5,285</td><td>277</td><td>4.3</td>

<td>120 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>120</td><td>5.311</td><td>279</td><td>6.1</td>

<td>120 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>120</td><td>5.311</td><td>278</td><td>6</td>

<td>140 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>145</td><td>5.329</td><td>X</td><td>12</td>

<td>140 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>145</td><td>5.329</td><td>309</td><td>11.7</td>

<td>160 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>168.7</td><td>5.343</td><td>X</td><td>18.8</td>

<td>160 mg / ml + 10 mM CaOAC + 6% sucrose</td><td>10</td><td>168.7</td><td>5.343</td><td>X</td><td>18.8</td>

[0117] The above-described experiments were performed for each of Ab-4, Ab-5, Ab-13, Ab14, Ab-19, Ab-20 and Ab-23 antibodies.

Example 3 - Effect of calcium acetate on other highly concentrated protein preparations [0118] The following example determined if calcium acetate reduces the viscosity of formulations containing high concentration of a protein other than the sclerostinic antibody.

[0119] The # 1 # 5 non-broweron antibodies were determined to have concentrations of 131.6 mg / ml, 94 mg / ml, 113.2 mg / ml, 50 mg / ml and 106.3, respectively. The term "non-kersterin antibody" as used herein, means an antibody other than the sclerostinic antibody described herein.

[0120] To 1 ml aliquots of 5 samples discussed above, 10 μl of 1.0 M Ca (OAc) was added₂. The viscosity, pH and osmolarity of the samples were assessed (see Table 6).

Table 6

<td>A sample</td><td>mg / ml</td><td>Viscosity (cP)</td>

<td>The non-kersteroste antibody # 1</td><td>94</td><td>6.8</td>

<td>The non-kersterostatin antibody # 1 + 10 mM Ca (OAc)₂</td><td>94</td><td>5.10</td>

<td>The non-kersteroste antibody # 2</td><td>135</td><td>9.8</td>

<td>The non-kersterostatin antibody # 2 + 10 mM Ca (OAc)₂</td><td>135</td><td>8.3</td>

<td>Protein # 1</td><td>50</td><td>3.3</td>

<td>Protein # 1 + 10 mM Ca (OAc)₂</td><td>50</td><td>3.2</td>

<td>Protein # 1</td><td>106.3</td><td>16.6</td>

<td>Protein # 1 + 10 mM Ca (OAc)₂</td><td>106.3</td><td>15.6</td>

[0121] Calcium acetate did not significantly reduce the viscosity of any of these samples.

Example 4 - Effect of calcium-free salt on the viscosity of a highly concentrated anti-sclerostin antibody preparation.

[0122] To determine if the calcium-derived salts will be able to reduce the viscosity of the anti-sclerostin antibody preparation, the following experiment was performed.

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[0123] The selected anti-sclerostin antibody (same as in Examples 1-2 above) was concentrated to ~ 130 mg / mL. To 1 ml of the antibody sample was added 10 μΐ 1.0 M (NH₄)₂SO₄ or 1.0 M MgSO₄. The viscosity of the control was set at 30 cP. It was found that MgSO4 significantly reduced the viscosity of the sample (MgSO4 + sample = 16 cP). (NH4) 2SO4 did not significantly reduce the viscosity of the sample.

Example 5 - Effect of other calcium salts on the viscosity of a highly concentrated anti-sclerostin antibody preparation [0124] To determine if calcium salts other than calcium acetate could reduce the viscosity of the sclerostin antibody preparation, the following experiment was performed.

[0125] The selected anti-sclerostin antibody (the same as in Examples 1-2 above) was concentrated to ~ 125 mg / ml. To 1 ml of the antibody sample was added 10 μl or 25 mM CaCl₂ or 25 mM MgCl₂. The viscosity of the control was set at 18.5 cP. It was found that CaCl2 and MgCl2 significantly reduced the viscosity of the sample (CaCl2 + sample = 9 cP and MgCl2 + sample = 8).

Example 6 - Effect of calcium acetate on another anti-sclerostin antibody [0126] To determine if calcium acetate would be able to reduce the viscosity of an anti-sclerostin antibody preparation containing an anti-sclerostin antibody other than in Examples 1-2 above, the following experiment was performed.

[0127] The selected anti-sclerostin antibody was concentrated to ~ 131 mg / ml. To 1 ml of the antibody sample was added 10 μl of 1.0 M Ca (OAc)₂. The control viscosity was determined to be 17.3 cP. It was established that Ca (OAc)₂ slightly decreased the viscosity of this sample (15.3 cP).

[0128] It is expected that those skilled in the art will be able to make numerous modifications and changes in the practice of the present invention to those skilled in the art when considered as being preferred. Accordingly, the only limitations to which the scope of the invention should be subject are those in the appended claims.

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SEQUENCE LIST [0129] <110> Timothy Osslund <120> Highly concentrated antibody preparations <130> 31173 / 45219A <140> US-61 / 334,986 <141> 2010-05-14 <160> 248 <170> Patentln version 3.5 < 210> 1 <211> 13 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-A and Ab-1 <400> 1

Gin Cheese Cheese Gin Cheese Val 1 5

Tyr Asp Asn Asn Trp Leu Ala 10 <210> 2 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-A and Ab-1 <400> 2

Asp Ala Cheese Asp Leu Ala Ser 1 5 <210> 3 <211> 10 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-A and Ab-1 <400> 3

Ala Tyr Asn Asp Val Ile Tyr 5

Gin Gly Ala Tyr Asn Asp Val Ile Tyr Ala 15 10 <210> 4 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-A and Ab-1 <400> 4

Cheese Tyr Trp Met Asn

5 <210> 5 <211> 16 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-1 and Ab-2 <400> 5

Asp Ser Gly Gly Arg Thr Asp Tyr 5 10

Thr 1 <210> 6 <211> 4 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-A and Ab-1 <400> 6

How much

Ala Ser Trp Ala

Lyi

Gly

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Asn Trp Asn Leu 1 <210> 7 <211> 654 <212> DNA <213> Artificial sequence <220>

<223> rabbit-mouse chimera <220>

<221> trait_different

<223> Light chain Ab-A (no signal) <400> 7

<td>gcgcaagtgc</td><td>tgacccagac</td><td>tccagcctcc</td><td>gtgtctgcag</td><td>ctgtgggagg</td><td>cacagtcacc</td><td>60</td>

<td>atcaattgcc</td><td>agtccagtca</td><td>gagtgtttat</td><td>gataacaact</td><td>ggttagcctg</td><td>gtttcagcag</td><td>120</td>

<td>aaaccagggc</td><td>agcctcccaa</td><td>gctcctgatt</td><td>tatgatgcat</td><td>ccgatctggc</td><td>atctggggtc</td><td>180</td>

<td>ccatcgcggt</td><td>tcagtggcag</td><td>tggatctggg</td><td>acacagttca</td><td>ctctcaccat</td><td>cagcggcgtg</td><td>240</td>

<td>cagtgtgccg</td><td>atgctgccac</td><td>ttactactgt</td><td>caaggcgctt</td><td>ataatgatgt</td><td>tatttatgct</td><td>300</td>

<td>ttcggcggag</td><td>ggaccgaggt</td><td>ggtggtcaaa</td><td>cgtacggatg</td><td>ctgcaccaac</td><td>tgtatccatc</td><td>360</td>

<td>ttcccaccat</td><td>ccagtgagca</td><td>gttaacatct</td><td>ggaggtgcct</td><td>cagtcgtgtg</td><td>cttcttgaac</td><td>420</td>

<td>aacttctacc</td><td>ccaaagacat</td><td>caatgtcaag</td><td>tggaagattg</td><td>atggcagtga</td><td>acgacaaaat</td><td>480</td>

<td>ggcgtcctga</td><td>acagttggac</td><td>tgatcaggac</td><td>agcaaagaca</td><td>gcacctacag</td><td>catgagcagc</td><td>540</td>

<td>accctcacgt</td><td>tgaccaagga</td><td>cgagtatgaa</td><td>cgacataaca</td><td>gctatacctg</td><td>tgaggccact</td><td>600</td>

<td>cacaagacat</td><td>caacttcacc</td><td>cattgtcaag</td><td>agcttcaaca</td><td>ggaatgagtg</td><td>ttag</td><td>654</td>

<210> 8 <211> 217 <212> PRT <213> Artificial sequence <220>

<223> rabbit-mouse chimera <220>

<221> CECHA_RÓŻN.

<223> Light chain Ab-A (no signal) <400> 8

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Ala Gin Val Leu Thr Gin Thr Pro Ala Ser 15 10

Val Ser Ala Ala Val Gly 15

Gly Thr Val Thr Ile Asn Cys Gin Cheese Ser 20 25

Gin Ser Val Tyr Asp Asn 30

Asn Trp Leu Ala Trp Phe Gin Gin Lys Pro 35 40

Gly Gin Pro Pro Lys Leu 45

Leu Ile Tyr Asp Ala Cheese Asp Leu Ala Ser 50 55

Gly Val Pro Ser Arg Phe 60

Cheese Gly Cheese Gly Cheese Gly Thr Gin Phe Thr 65 70

Leu Thr Ile Ser Gly Val 75 80

Gin Cys Ala Asp Ala Ala Thr Tyr Tyr Cys 85 90

Gin Gly Ala Tyr Asn Asp 95

Val Ile Tyr Ala Phe Gly Gly Gly Thr Glu 100 105

Val Val Val Lys Arg Thr 110

Asp Ala Ala Pro Thr Val Ser Ile Phe Pro 115 120

Pro Ser Ser Glu Gin Leu 125

Thr Ser Gly Gly Ala Ser Val Val Cys Phe 130 135

Leu Asn Asn Phe Tyr Pro 140

Lys Asp. Ile Asn. Val Lys Trp. Lys. Ile Asp 145. 150

Gly Ser Glu Arg Gin Asn 155 160

Gly Val Leu Asn Cheese Trp Thr Asp Gin Asp 165 170

Lys Asp Ser Thr Tyr 175

Ser Met Ser Ser Thr Leu Thr Leu Thr Lys 180 185

Asp Glu Tyr Glu Arg His 190

Asn Cheese Tyr Thr Cys Glu Ala Thr His Lys 195 200

Thr Ser Thr Cheese Pro Ile 205

Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 9 <211> 1302 <212> DNA <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> trait_different

<223> Ab-A heavy chain (no signal) <400> 9

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EP 2 569 010 B1 cagtcgctgg aggagtccgg gggtcgcctg gtcacgcctg ggacacccct gacactcacc 60 tgcacagcct ctggattctc cctcagtagt tattggatga actgggtccg ccaggctcca 120 ggggaggggc tggaatggat cggaaccatt gattctggtg gtaggacgga ctacgcgagc 180 tgggcaaaag gccgattcac catctccaga acctcgacta cgatggatct gaaaatgacc 240 agtctgacga ccggggacac ggcccgttat ttctgtgcca gaaattggaa cttgtggggc 300 caaggcaccc tcgtcaccgt ctcgagcgct tctacaaagg gcccatctgt ctatccactg 360 gcccctggat ctgctgccca aactaactcc atggtgaccc tgggatgcct ggtcaagggc 420 tatttccctg agccagtgac agtgacctgg aactctggat ccctgtccag cggtgtgcac 480 accttcccag ctgtcctgca gtctgacctc tacactctga gcagctcagt gactgtcccc 540 tccagcacct ggcccagcga gaccgtcacc tgcaacgttg cccacccggc cagcagcacc 600 aaggtggaca agaaaattgt gcccagggat tgtggttgta agccttgcat atgtacagtc 660 ccagaagtat catctgtctt catcttcccc ccaaagccca aggatgtgct caccattact 720ctgactccta aggtcacgtg tgttgtggta gacatcagca aggatgatcc cgaggtccag 780 ttcagctggt ttgtagatga tgtggaggtg cacacagctc agacgcaacc ccgggaggag 840 cagttcaaca gcactttccg ctcagtcagt gaacttccca tcatgcacca ggactggctc 900 aatggcaagg agttcaaatg cagggtcaac agtgcagctt tccctgcccc catcgagaaa 960 accatctcca aaaccaaagg cagaccgaag gctccacagg tgtacaccat tccacctccc 1020 aaggagcaga tggccaagga taaagtcagt ctgacctgca tgataacaga cttcttccct 1080 gaagacatta ctgtggagtg gcagtggaat gggcagccag cggagaacta caagaacact 1140 cagcccatca tggacacaga tggctcttac ttcgtctaca gcaagctcaa tgtgcagaag 1200 agcaactggg aggcaggaaa tactttcacc tgctctgtgt tacatgaggg cctgcacaac 1260 caccatactg agaagagcct ctcccactct cctggtaaat ga 1302 <210> 10 <211> 433 <212> PRT <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> CECHA_RÓŻN.

<223> Ab-A heavy chain (no signal) <400> 10

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Gin Cheese Leu Glu Glu Cheese Gly Gly Arg Leu Val Thr Pro Gly Thr Pro 15 10 15

Leu Thr Leu Thr Cys Thr Ala Cheese Gly Phe Cheese Leu Cheese Cheese Tyr Trp 20 25 30

Met Asn Trp Val Arg Gin Ala Pro Gly Glu Gly Leu Glu Trp Ile Gly 35 40 45

Thr Ile Asp Ser Gly Gly Arg Thr Asp Tyr Ala Ser Trp Ala Lys Gly 50 55 60

Arg Phe Thr Ile Ser Arg Thr Ser Thr Thr Met Asp Leu Ly3 Met Thr 65 70 75 80

Leu Thr Thr Thr Gly Asp Thr Ala Arg Tyr Phe Cys Ala Arg Asn Trp

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90 95

Asn Leu Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 100 105 110

Lys Gly Pro Cheese Val Tyr Pro Leu Ala Pro Gly Cheese Ala Ala Gin Thr 115 120 125

Asn Cheese Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 130 135 140

Pro Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His 145 150 155 160

Thr Phe Pro Ala Val Leu Gin Le Asp cheese Leu Tyr Thr Leu Cheese Ser Ser 165 Π0 175

Val Thr Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn 180 185 190

Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro 195 200 205

Arg Asp Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser 210 215 220

Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr 225 230 235 240

Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Ile Ile Lys Asp Asp 245 250 255

Pro Glu Val Gin Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr 260 265 270

Ala Gin Thr Gin Pro Arg Glu Gin Phe Asn Ser Thr Phe Arg Ser 275 280 285

Val Ser Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu 290 295 300

Phe Lys Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys 305 310 315 320

Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys Al Pro Gin Val Tyr Thr

VP / 4572 / AGR

EP 2 569 010 B1

325 330 335

How much Pro Pro Pro Lys Glu Gin Met Al Lys Asp Lys Val Ser Leu Thr 340 345 350

Cys Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin 355 360 365

Trp Asn Gly Gin Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro Ile Met 370 375 380

Asn Thr Asn Gly Cheese Tyr Phe Val Tyr Cheese Lys Leu Asn Val Gin Lys 385 390 395 400

Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu 405 410 415

Gly Leu His Asn His Thr Glu Lys Ser Leu Cheese His Cheese Pro Gly 420 425 430

Lys <210> 11 <211> 390 <212> DNA <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> trait_different

<223> Lightweight variable Ab-1 (with signal) <400> 11

<td>atggacacga</td><td>gggcccccac</td><td>tcagctgctg</td><td>gggctcctgc</td><td>tgctctggct</td><td>cccaggtgcc</td><td>60</td>

<td>acatttgctc</td><td>aagttctgac</td><td>ccagagtcca</td><td>agcagtctct</td><td>ccgccagcgt</td><td>aggcgatcgt</td><td>120</td>

<td>gtgactatta</td><td>cctgtcaatc</td><td>tagtcagagc</td><td>gtgtatgata</td><td>acaattggct</td><td>ggcgtggtac</td><td>180</td>

<td>cagcaaaaac</td><td>cgggcaaagc</td><td>cccgaagctg</td><td>ctcatctatg</td><td>acgcgtccga</td><td>tctggctagc</td><td>240</td>

<td>ggtgtgccaa</td><td>gccgtttcag</td><td>tggcagtggc</td><td>agcggtactg</td><td>actttaccct</td><td>cacaatttcg</td><td>300</td>

<td>tctctccagc</td><td>cggaagattt</td><td>cgccacttac</td><td>tattgtcaag</td><td>gtgcttacaa</td><td>cgatgtgatt</td><td>360</td>

<td>tatgccttcg</td><td>gtcagggcac</td><td>taaagtagaa</td><td></td><td></td><td></td><td>390</td>

<210> 12 <211> 133 <212> PRT <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> CECHA_RÓŻN.

<223> Lightweight variable Ab-1 (with signal) <400> 12

VP / 4572 / AGR

EP 2 569 010 B1

Met Asp Thr Arg Ala Pro Thr Gin Leu Leu Gly Leu Leu Leu Trp 15 10 15

Leu Pro Gly Ala Thr Phe Ala Gin Val Leu Thr Gin Cheese Pro Cheese Ser 20 25 30

Leu Cheese Ala Cheese Val Gly Asp Arg Val Thr Ile Thr Cys Gin Cheese Ser 35 40 45

Gin Ser Val Tyr Asp Asn Asn Trp Leu Ala Trp Tyr Gin Gin Lys Pro 50 55 60

Gly Lys Ala Pro Lys Leu Leu Ile Tyr Asp Ala Cheese Asp Leu Ala Ser 65 70 75 80

Gly Val Pro Ser Arg Phe Cheese Gly Cheese Gly Cheese Gly Thr Asp Phe Thr 85 90 95

Leu Thr Ile Ser Ser Leu Gin Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 100 105 110

Gin Gly Ala Tyr Asn Asp Val Ile Tyr Ala Phe Gly Gin Gly Thr Lys 115 120 125

Val Glu Ile Lys Arg 130 <210> 13 <211> 393 <212> DNA <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> trait_different

<223> Heavy variable Ab-1 (with signal) <400> 13

<td>atggagactg</td><td>ggctgcgctg</td><td>gcttctcctg</td><td>gtcgctgtgc</td><td>tcaaaggtgt</td><td>ccactgtgag</td><td>60</td>

<td>gtgcagctgt</td><td>tggagtctgg</td><td>aggcgggctt</td><td>gtccagcctg</td><td>gagggagcct</td><td>gcgtctctct</td><td>120</td>

<td>tgtgcagcaa</td><td>gcggcttcag</td><td>cttatcctct</td><td>tactggatga</td><td>attgggtgcg</td><td>gcaggcacct</td><td>180</td>

<td>gggaagggcc</td><td>tggagtgggt</td><td>gggcaccatt</td><td>gattccggag</td><td>gccgtacaga</td><td>ctacgcgtct</td><td>240</td>

<td>tgggcaaagg</td><td>gccgtttcac</td><td>catttcccgc</td><td>gacaactcca</td><td>aaaataccat</td><td>gtacctccag</td><td>300</td>

<td>atgaactctc</td><td>tccgcgcaga</td><td>ggacacagca</td><td>cgttattact</td><td>gtgcacgcaa</td><td>ctggaatctg</td><td>360</td>

<td>tggggtcaag</td><td>gtactcttgt</td><td>aacagtctcg</td><td>agc</td><td></td><td></td><td>393</td>

<210> 14 <211> 131 <212> PRT <213> Artificial sequence <220>

<223> Humanized antibody <400> 14

VP / 4572 / AGR

EP 2 569 010 B1

Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly 15 10 15

Val His Cys Glu Val Gin Leu Leu Glu Gly Gly Gly Leu Val Gin 20 25 30

Pro Gly Gly Leu Arg Leu Cheese Ser Cys Ala Ala Ser Gly Phe Leu 35 35 45

Cheese Cheese Tyr Trp Met Asn Trp Val Arg Gin Al Pro Gly Lys Gly Leu 50 55 60

Glu Trp Val Gly Thr Ile Asp Ser Gly Gly Arg Thr Asp Tyr Ala Ser 65 70 75 80

Trp Ala Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Ly3 Asn Thr 85 90 95

Met Tyr Leu Gin Met Asn Leu Ser Arg Ala Glu Asp Thr Ala Arg Tyr 100 105 110

Tyr Cys Ala Arg Asn Trp A3n Leu Trp Gly Gin Gly Thr Leu Val Thr 115 120 125

Val Ser Ser 130 <210> 15 <211> 10 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-B <400> 15

Ser Ala Ser Ser Ser Ser Val Ser Phe Val Asp 15 10 <210> 16 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-B <400> 16

Arg Thr Ser Asn Leu Gly Phe 1 5 <210> 17 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 <400> 17

Gin Gin Arg Ser Thr Tyr Pro Pro Thr 1 5 <210> 18 <211> 7

VP / 4572 / AGR

EP 2 569 010 B1 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-B <400> 18

Thr Ser Gly Met Gly Val Gly 1 5 <210> 19 <211> 16 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-B <400> 19

His Asp. Asp Asp Val Val Lys Arg Tyr Asn Pro Val Leu Lys Ser 15 10 15 <210> 20 <211> 14 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-B <400> 20

Glu Asp Phe Asp Tyr Asp Glu Glu Tyr Tyr Ala Met Asp Tyr 15 10 <210> 21 <211> 642 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-B light chain (no signal) <400> 21

<td>caaattgttc</td><td>tcacccagtc</td><td>tccaacaatc</td><td>gtgtctgcat</td><td>ctccagggga</td><td>gaaggtcacc</td><td>60</td>

<td>ctaatctgca</td><td>gtgccagttc</td><td>aagtgtaagt</td><td>ttcgtggact</td><td>ggttccagca</td><td>gaagccaggc</td><td>120</td>

<td>acttctccca</td><td>aacgctggat</td><td>ttacagaaca</td><td>tccaacctgg</td><td>gttttggagt</td><td>ccctgctcgc</td><td>180</td>

<td>ttcagtggcg</td><td>gtggatctgg</td><td>gacctctcac</td><td>tctctcacaa</td><td>tcagccgaat</td><td>ggaggctgaa</td><td>240</td>

<td>gatgctgcca</td><td>cttattactg</td><td>ccagcaaagg</td><td>agtacttacc</td><td>cacccacgtt</td><td>cggtgctggg</td><td>300</td>

<td>accaagctgg</td><td>aactgaaacg</td><td>ggctgatgct</td><td>gcaccaactg</td><td>tatccatctt</td><td>cccaccatcc</td><td>360</td>

<td>agtgagcagt</td><td>taacatctgg</td><td>aggtgcctca</td><td>gtcgtgtgct</td><td>tcttgaacaa</td><td>cttctacccc</td><td>420</td>

<td>aaagacatca</td><td>atgtcaagtg</td><td>gaagattgat</td><td>ggcagtgaac</td><td>gacaaaatgg</td><td>cgtcctgaac</td><td>480</td>

<td>agttggactg</td><td>atcaggacag</td><td>caaagacagc</td><td>acctacagca</td><td>tgagcagcac</td><td>cctcacgttg</td><td>540</td>

<td>accaaggacg</td><td>agtatgaacg</td><td>acataacagc</td><td>tatacctgtg</td><td>aggccactca</td><td>caagacatca</td><td>600</td>

<td>acttcaccca</td><td>ttgtcaagag</td><td>cttcaacagg</td><td>aatgagtgtt</td><td>ag</td><td></td><td>642</td>

<210> 22 <211> 213 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-B light chain (no signal) <400> 22

VP / 4572 / AGR

EP 2 569 010 B1

Gin Ile Val Leu Thr Gin Cheese Pro Thr 1 5

Glu Lys Val Thr Leu Ile Cys Ser Ala 20 25

Asp Trp Phe Gin Gin Lys Pro Gly Thr 35 40

Arg Thr Ser A3n Leu Gly Phe Gly Val 50 55

Gly Cheese Gly Thr Cheese His Cheese Leu Thr 65 70

Asp Ala Ala Thr Tyr Tyr Cys Gin Gin 85

Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105

Thr Val Ser Ile Phe Pro Pro Ser Ser 115 120

Ala Ser Val Val Cys Phe Leu Asn Asn 130 135

Val Lys Trp Ly3 Ile Asp Gly Cheese Glu 145 150

Trp Asp Asp. Gin Asp Ser Lys Asp 165

Thr Leu Thr Leu Thr Lys Asp Glu Tyr 180 185

Cys Glu Ala Thr His Lys Thr Ser Thr 195 200

How much

Cheese

Cheese

Pro

How much

Arg

lys

Glu

phe

Arg

Cheese

170

Glu

Cheese

Val Ser Ala Ser Pro Gly 15

Ser Ser Val Ser Phe Val 30

Pro Lys Arg Trp Ile Tyr 45

Ala Arg Phe Cheese Gly Gly 60

Ser Arg Met Glu Ala Glu 75 80

Thr Tyr Pro Pro Thr 95 cheese

Arg Ala Asp Ala Ala Pro 110

Gin Leu Thr Cheese Gly Gly 125

Tyr Pro Lys Asp Asn 140

Gin Asn Gly Val Leu Asn 155 160

Thr Tyr Cheese Met Ser Ser 175

Arg His Asn Cheese Tyr Thr 190

Pro Ile Val Lys Ser Phe 205

Asn Arg Asn Glu Cys 210 <210> 23 <211> 1350 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-B heavy chain (no signal) <400> 23

VP / 4572 / AGR

EP 2 569 010 B1

<td>caggttactc</td><td>tgaaagagtc</td><td>tggccctggg</td><td>atattgcagc</td><td>cctcccagac</td><td>cctcagtctg</td><td>60</td>

<td>acttgttctt</td><td>tctctgggtt</td><td>ttcactgagc</td><td>acttctggta</td><td>tgggtgtagg</td><td>ctggattcgt</td><td>120</td>

<td>cacccatcag</td><td>ggaagaatct</td><td>ggagtggctg</td><td>gcacacattt</td><td>ggtgggatga</td><td>tgtcaagcgc</td><td>180</td>

<td>tataacccag</td><td>tcctgaagag</td><td>ccgactgact</td><td>atctccaagg</td><td>atacctccaa</td><td>cagccaggta</td><td>240</td>

<td>ttcctcaaga</td><td>tcgccaatgt</td><td>ggacactgca</td><td>gatactgcca</td><td>catactactg</td><td>tgctcgaata</td><td>300</td>

<td>gaggactttg</td><td>attacgacga</td><td>ggagtattat</td><td>gctatggact</td><td>actggggtca</td><td>aggaacctca</td><td>360</td>

<td>gtcatcgtct</td><td>cctcagccaa</td><td>aacgacaccc</td><td>ccatctgtct</td><td>atccactggc</td><td>ccctggatct</td><td>420</td>

<td>gctgcccaaa</td><td>ctaactccat</td><td>ggtgaccctg</td><td>ggatgcctgg</td><td>tcaagggcta</td><td>tttccctgag</td><td>480</td>

<td>ccagtgacag</td><td>tgacctggaa</td><td>ctctggatcc</td><td>ctgtccagcg</td><td>gtgtgcacac</td><td>cttcccagct</td><td>540</td>

<td>gtcctgcagt</td><td>ctgacctcta</td><td>cactctgagc</td><td>agctcagtga</td><td>ctgtcccctc</td><td>cagcacctgg</td><td>600</td>

<td>cccagcgaga</td><td>ccgtcacctg</td><td>caacgttgcc</td><td>cacccggcca</td><td>gcagcaccaa</td><td>ggtggacaag</td><td>660</td>

<td>aaaattgtgc</td><td>ccagggattg</td><td>tggttgtaag</td><td>ccttgcatat</td><td>gtacagtccc</td><td>agaagtatca</td><td>720</td>

<td>tctgtcttca</td><td>tcttcccccc</td><td>aaagcccaag</td><td>gatgtgctca</td><td>ccattactct</td><td>gactcctaag</td><td>780</td>

<td>gtcacgtgtg</td><td>ttgtggtaga</td><td>catcagcaag</td><td>gatgatcccg</td><td>aggtccagtt</td><td>cagctggttt</td><td>840</td>

<td>gtagatgatg</td><td>tggaggtgca</td><td>cacagctcag</td><td>acgcaacccc</td><td>gggaggagca</td><td>gttcaacagc</td><td>900</td>

<td>actttccgct</td><td>cagtcagtga</td><td>acttcccatc</td><td>atgcaccagg</td><td>actggctcaa</td><td>tggcaaggag</td><td>960</td>

<td>ttcaaatgca</td><td>gggtcaacag</td><td>tgcagctttc</td><td>cctgccccca</td><td>tcgagaaaac</td><td>catctccaaa</td><td>1020</td>

<td>accaaaggca</td><td>gaccgaaggc</td><td>tccacaggtg</td><td>tacaccattc</td><td>cacctcccaa</td><td>ggagcagatg</td><td>1080</td>

<td>gccaaggata</td><td>aagtcagtct</td><td>gacctgcatg</td><td>ataacagact</td><td>tcttccctga</td><td>agacattact</td><td>1140</td>

<td>gtggagtggc</td><td>agtggaatgg</td><td>gcagccagcg</td><td>gagaactaca</td><td>agaacactca</td><td>gcccatcatg</td><td>1200</td>

<td>gacacagatg</td><td>gctcttactt</td><td>cgtctacagc</td><td>aagctcaatg</td><td>tgcagaagag</td><td>caactgggag</td><td>1260</td>

<td>gcaggaaata</td><td>ctttcacctg</td><td>ctctgtgtta</td><td>catgagggcc</td><td>tgcacaacca</td><td>ccatactgag</td><td>1320</td>

aagagcctct cccactctcc tggtaaatga 1350 <210> 24 <211> 449 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-B heavy chain (no signal) <400> 24

VP / 4572 / AGR

EP 2 569 010 B1

Gin Val Thr Leu Lys Glu Cheese Gly Pro Gly Ile Leu Gin Pro Cheese Gin 15 10 15

Thr Leu Cheese Leu Thr Cys Cheese Phe Cheese Gly Phe Cheese Leu Cheese Thr Ser 20 25 30

Gly Met Gly Val Gly Trp Ile Arg His Pro Ser Gly Lys Asn Leu Glu 35 40 45

Trp Leu Ala His Trp Trp Asp Asp Val Lys Arg Tyr Asn Pro Val 50 55 60

Leu Lys Ser Arg Leu Thr Ile Cheese Lys Asp Thr Cheese Asn Cheese Gin Val 65 70 75 80

Phe Leu Lys Ile Ala Asn Val Asp Thr Ala Asp Thr Ala Thr Tyr Tyr 85 90 95

Cys Ala Arg Ile Glu Asp Asp. Asp. Asp Glu Glu Tyr Tyr Ala Met 100 105 110

Asp Tyr Trp Gly Gin Gly Thr Ser Val Ile Val Ser Ser Al Lys Thr 115 120 125

Thr Pro Pro Ser Val Tire Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr 130 135 140

Asn Cheese Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 145 150 155 160

Pro Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His 165 170 175

Phe Thr Pro Ala Val Leu Leu Gln Asp Ser Tyr Leu Thr cheese Se r

VP / 4572 / AGR

EP 2 569 010 B1

180 185 190

Val Thr Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn 195 200 205

Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro 210 215 220

Arg Asp Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser 225 230 235 240

Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr 245 250 255

Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Ile Ile Lys Asp Asp 260 265 270

Pro Glu Val Gin Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr 275 280 285

Ala Gin Thr Gin Pro Arg Glu Gin Phe Asn Ser Thr Phe Arg Cheese 290 295 300

Val Ser Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Phe Lys Cys Arg val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys 325 330 335

Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys Al Pro Gin Val Tyr Thr 340 345 350

How much Pro Pro Pro Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr 355 360 365

Cys Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin 370 375 380

Trp Asn Gly Gin Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro Ile Met 385 390 395 400

Asp Thr Asp Gly Cheese Tyr Phe Val Tyr Cheese Lys Leu Asn Val Gin Lys 405 410 415

Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu

VP / 4572 / AGR

EP 2 569 010 B1

<img file="PL2569010T3_D0001.tif" />

<210> 25 <211> 15 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-C <400> 25

Cheese Gin Cheese Val Asp Tyr Asp 5

ala

Lys 1 <210> 26 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-C <400> 26

Ala Ala Ser Asn Leu Glu Ser 1 5 <210> 27 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-C <400> 27

Asn Glu Asp Pro Trp Thr 5

Gin

Gin 1 <210> 28 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-C <400> 28

Asp Cys Tyr Met Asn

5 <210> 29 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-C <400> 29

Asp Asn Pro Phe Asn Gly Gly Thr 1 5

Gly <210> 30 <211> 16 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-C

Gly

Thr

Asp Ser Tyr Met

Tyr Asn Gin Lys

own

phe

lys

VP / 4572 / AGR

EP 2 569 010 B1 &amp; lt; 400 &amp; gt; 30

Ser His Tyr Tyr Phe Asp Gly Arg Val Pro Trp Asp Ala Met Asp Tyr 15 10 15 <210> 31 <211> 657 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-C light chain (no signal) <400> 31

<td>gacattgtgc</td><td>tgacccaatc</td><td>tccagcttct</td><td>ttgactgtgt</td><td>ctctaggcct</td><td>gagggccacc</td><td>60</td>

<td>atctcctgca</td><td>aggccagcca</td><td>aagtgttgat</td><td>tatgatggtg</td><td>atagttatat</td><td>gaactggtac</td><td>120</td>

<td>cagcagaaac</td><td>caggacagcc</td><td>acccaaactc</td><td>ctcatctatg</td><td>ctgcatccaa</td><td>tctagaatct</td><td>180</td>

<td>gggatcccag</td><td>ccaggtttag</td><td>tggcaatggg</td><td>tctgggacag</td><td>acttcaccct</td><td>caacatccat</td><td>240</td>

<td>cctgtggagg</td><td>aggaggatgc</td><td>tgtaacctat</td><td>tactgtcaac</td><td>aaagtaatga</td><td>ggatccgtgg</td><td>300</td>

<td>acgttcggtg</td><td>gaggcaccaa</td><td>gctggaaatc</td><td>aaacgggctg</td><td>atgctgcacc</td><td>aactgtatcc</td><td>360</td>

<td>atcttcccac</td><td>catccagtga</td><td>gcagttaaca</td><td>tctggaggtg</td><td>cctcagtcgt</td><td>gtgcttcttg</td><td>420</td>

<td>aacaacttct</td><td>accccaaaga</td><td>catcaatgtc</td><td>aagtggaaga</td><td>ttgatggcag</td><td>tgaacgacaa</td><td>480</td>

<td>aatggcgtcc</td><td>tgaacagttg</td><td>gactgatcag</td><td>gacagcaaag</td><td>acagcaccta</td><td>cagcatgagc</td><td>540</td>

<td>agcaccctca</td><td>cgttgaccaa</td><td>ggacgagtat</td><td>gaacgacata</td><td>acagctatac</td><td>ctgtgaggcc</td><td>600</td>

<td>actcacaaga</td><td>catcaacttc</td><td>acccattgtc</td><td>aagagcttca</td><td>acaggaatga</td><td>gtgttag</td><td>657</td>

<210> 32 <211> 218 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-C light chain (no signal) <400> 32

EP 2 569 010 B1

<td>VP / 4572</td><td colspan="2">/ AGR</td><td></td><td></td><td></td><td></td><td></td><td></td><td colspan="2">47</td><td></td><td></td><td></td><td></td><td></td>

<td>Asp</td><td>How much</td><td>val</td><td>Leu</td><td>Thr</td><td>Gin</td><td>Cheese</td><td>Pro</td><td>ala</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>val</td><td>Cheese</td><td>Leu</td><td>Gly</td>

<td>1</td><td></td><td></td><td></td><td>5</td><td></td><td></td><td></td><td></td><td>10</td><td></td><td></td><td></td><td></td><td>15</td><td></td>

<td>Leu</td><td>Arg</td><td>ala</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cys</td><td>lys</td><td>ala</td><td>Cheese</td><td>Gin</td><td>Cheese</td><td>val</td><td>Asp</td><td>Tyr</td><td>Asp</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>Gly</td><td>Asp</td><td>Cheese</td><td>Tyr</td><td>Underworld</td><td>own</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Ly3</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Pro</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>lys</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td><td>ala</td><td>ala</td><td>Cheese</td><td>own</td><td>Leu</td><td>Glu</td><td>Cheese</td><td>Gly</td><td>How much</td><td>Pro</td><td>ala</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Arg</td><td>phe</td><td>Cheese</td><td>Gly</td><td>own</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>phe</td><td>Thr</td><td>Leu</td><td>own</td><td>How much</td><td>His</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Pro</td><td>val</td><td>Glu</td><td>Glu</td><td>Glu</td><td>A3P</td><td>ala</td><td>val</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Cheese</td><td>own</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>Glu</td><td>Asp</td><td>Pro</td><td>Trp</td><td>Thr</td><td>phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>lys</td><td>Leu</td><td>Glu</td><td>How much</td><td>lys</td><td>Arg</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>ala</td><td>Asp</td><td>ala</td><td>ala</td><td>Pro</td><td>Thr</td><td>val</td><td>Cheese</td><td>How much</td><td>phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Cheese</td><td>Glu</td><td>Gin</td>

<td></td><td></td><td>115</td><td></td><td></td><td></td><td></td><td>120</td><td></td><td></td><td></td><td></td><td>125</td><td></td><td></td><td></td>

<td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>ala</td><td>Cheese</td><td>val</td><td>val</td><td>Cys</td><td>phe</td><td>Leu</td><td>Α3Π</td><td>own</td><td>phe</td><td>Tyr</td>

<td></td><td>130</td><td></td><td></td><td></td><td></td><td>135</td><td></td><td></td><td></td><td></td><td>140</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Ly3</td><td>Asp</td><td>How much</td><td>own</td><td>val</td><td>Ly3</td><td>Trp</td><td>lys</td><td>How much</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Glu</td><td>Arg</td><td>Gin</td>

<td>145</td><td></td><td></td><td></td><td></td><td>150</td><td></td><td></td><td></td><td></td><td>155</td><td></td><td></td><td></td><td></td><td>160</td>

<td>own</td><td>Gly</td><td>val</td><td>Leu</td><td>own</td><td>Cheese</td><td>Trp</td><td>Thr</td><td>Asp</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>lys</td><td>Asp</td><td>Cheese</td><td>Thr</td>

<td></td><td></td><td></td><td></td><td>165</td><td></td><td></td><td></td><td></td><td>170</td><td></td><td></td><td></td><td></td><td>175</td><td></td>

<td>Tyr</td><td>Cheese</td><td>Underworld</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Thr</td><td>lys</td><td>Asp</td><td>Glu</td><td>Tyr</td><td>Glu</td><td>Arg</td>

<td></td><td></td><td></td><td>180</td><td></td><td></td><td></td><td></td><td>185</td><td></td><td></td><td></td><td></td><td>190</td><td></td><td></td>

<td>His</td><td>own</td><td>Cheese</td><td>Tyr</td><td>Thr</td><td>Cys</td><td>Glu</td><td>ala</td><td>Thr</td><td>His</td><td>lys</td><td>Thr</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Pro</td>

<td></td><td></td><td>195</td><td></td><td></td><td></td><td></td><td>200</td><td></td><td></td><td></td><td></td><td>205</td><td></td><td></td><td></td>

<td>How much</td><td>val</td><td>lys</td><td>Cheese</td><td>phe</td><td>own</td><td>Arg</td><td>own</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>210</td><td></td><td></td><td></td><td></td><td>215</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<210> 33 <211> 1350 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-C heavy chain (no signal) <400> 33

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggtccagc</td><td>tgcaacaatc</td><td>tggacctgag</td><td>ctggtgaagc</td><td>ctgggacttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgtaagg</td><td>cttctggata</td><td>cacattcact</td><td>gactgctaca</td><td>tgaactgggt</td><td>gaagcagagc</td><td>120</td>

<td>catgggaaga</td><td>gccttgaatg</td><td>gattggagat</td><td>attaatcctt</td><td>tcaacggtgg</td><td>tactacctac</td><td>180</td>

<td>aaccagaagt</td><td>tcaagggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>aatcctccag</td><td>cacagcctac</td><td>240</td>

<td>atgcagctca</td><td>acagcctgac</td><td>atctgacgac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagatcccat</td><td>300</td>

<td>tattacttcg</td><td>atggtagagt</td><td>cccttgggat</td><td>gctatggact</td><td>actggggtca</td><td>aggaacctca</td><td>360</td>

<td>gtcaccgtct</td><td>cctcagccaa</td><td>aacgacaccc</td><td>ccatctgtct</td><td>atccactggc</td><td>ccctggatct</td><td>420</td>

<td>gctgcccaaa</td><td>ctaactccat</td><td>ggtgaccctg</td><td>ggatgcctgg</td><td>tcaagggcta</td><td>tttccctgag</td><td>480</td>

<td>ccagtgacag</td><td>tgacctggaa</td><td>ctctggatcc</td><td>ctgtccagcg</td><td>gtgtgcacac</td><td>cttcccagct</td><td>540</td>

<td>gtcctgcagt</td><td>ctgacctcta</td><td>cactctgagc</td><td>agctcagtga</td><td>ctgtcccctc</td><td>cagcacctgg</td><td>600</td>

<td>cccagcgaga</td><td>ccgtcacctg</td><td>caacgttgcc</td><td>cacccggcca</td><td>gcagcaccaa</td><td>ggtggacaag</td><td>660</td>

<td>aaaattgtgc</td><td>ccagggattg</td><td>tggttgtaag</td><td>ccttgcatat</td><td>gtacagtccc</td><td>agaagtatca</td><td>720</td>

<td>tctgtcttca</td><td>tcttcccccc</td><td>aaagcccaag</td><td>gatgtgctca</td><td>ccattactct</td><td>gactcctaag</td><td>780</td>

<td>gtcacgtgtg</td><td>ttgtggtaga</td><td>catcagcaag</td><td>gatgatcccg</td><td>aggtccagtt</td><td>cagctggttt</td><td>840</td>

<td>gtagatgatg</td><td>tggaggtgca</td><td>cacagctcag</td><td>acgcaacccc</td><td>gggaggagca</td><td>gttcaacagc</td><td>900</td>

<td>actttccgct</td><td>cagtcagtga</td><td>acttcccatc</td><td>atgcaccagg</td><td>actggctcaa</td><td>tggcaaggag</td><td>960</td>

<td>ttcaaatgca</td><td>gggtcaacag</td><td>tgcagctttc</td><td>cctgccccca</td><td>tcgagaaaac</td><td>catctccaaa</td><td>1020</td>

<td>accaaaggca</td><td>gaccgaaggc</td><td>tccacaggtg</td><td>tacaccattc</td><td>cacctcccaa</td><td>ggagcagatg</td><td>1080</td>

<td>gccaaggata</td><td>aagtcagtct</td><td>gacctgcatg</td><td>ataacagact</td><td>tcttccctga</td><td>agacattact</td><td>1140</td>

<td>gtggagtggc</td><td>agtggaatgg</td><td>gcagccagcg</td><td>gagaactaca</td><td>agaacactca</td><td>gcccatcatg</td><td>1200</td>

<td>gacacagatg</td><td>gctcttactt</td><td>catctacagc</td><td>aagctcaatg</td><td>tgcagaagag</td><td>caactgggag</td><td>1260</td>

<td>gcaggaaata</td><td>ctttcacctg</td><td>ctctgtgtta</td><td>catgagggcc</td><td>tgcacaacca</td><td>ccatactgag</td><td>1320</td>

<td>aagagcctct</td><td>cccactctcc</td><td>tggtaaatga</td><td></td><td></td><td></td><td>1350</td>

<210> 34 <211> 449 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-C heavy chain (no signal) <400> 34

PZ / 4572 / AGR 49

EP 2 569 010 B1

<td rowspan="2">Glu 1</td><td rowspan="2">val</td><td colspan="2" rowspan="2">Gin Leu</td><td colspan="8">Gin Gin Cheese Gly Pro Glu Leu Val</td><td rowspan="2">Ly3</td><td rowspan="2">Pro</td><td rowspan="2">Gly 15</td><td rowspan="2">Thr</td>

<td colspan="2">5</td><td colspan="6">10</td>

<td>Cheese</td><td>val</td><td>Lye</td><td>Underworld</td><td>Cheese</td><td>Cys</td><td>lys</td><td>ala</td><td>Cheese</td><td>Gly</td><td>Tyr</td><td>Thr</td><td>phe</td><td>Thr</td><td>Asp</td><td>Cys</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>Tyr</td><td>Underworld</td><td>own</td><td>Trp</td><td>val</td><td>lys</td><td>Gin</td><td>Cheese</td><td>His</td><td>Gly</td><td>lys</td><td>Cheese</td><td>Leu</td><td>Glu</td><td>Trp</td><td>How much</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Gly</td><td>Asp</td><td>How much</td><td>Α3Π</td><td>Pro</td><td>phe</td><td>own</td><td>Gly</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Tyr</td><td>own</td><td>Gin</td><td>lys</td><td>phe</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>lys</td><td>Gly</td><td>lys</td><td>ala</td><td>Thr</td><td>Leu</td><td>Thr</td><td>val</td><td>Asp</td><td>lys</td><td>Cheese</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>ala</td><td>Tyr</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Underworld</td><td>Gin</td><td>Leu</td><td>Α3Π</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Asp</td><td>Asp</td><td>Cheese</td><td>ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cye</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>ala</td><td>Arg</td><td>Cheese</td><td>His</td><td>Tyr</td><td>Tyr</td><td>phe</td><td>Asp</td><td>Gly</td><td>Arg</td><td>val</td><td>Pro</td><td>Trp</td><td>Asp</td><td>ala</td><td>Underworld</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Thr</td><td>Cheese</td><td>val</td><td>Thr</td><td>val</td><td>Cheese</td><td>Cheese</td><td>ala</td><td>Ly3</td><td>Thr</td>

<td></td><td></td><td>115</td><td></td><td></td><td></td><td></td><td>120</td><td></td><td></td><td></td><td></td><td>125</td><td></td><td></td><td></td>

<td>Thr</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>val</td><td>Tyr</td><td>Pro</td><td>Leu</td><td>ala</td><td>Pro</td><td>Gly</td><td>Cheese</td><td>ala</td><td>ala</td><td>Gin</td><td>Thr</td>

VP / 4572 / AGR

EP 2 569 010 B1

130 135 140

Asn Cheese Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 145 150 155 160

Pro Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His 165 170 175

Thr Phe Pro Ala Val Leu Gin Le Asp cheese Leu Tyr Thr Leu Ser Ser Ser 180 185 190

Val Thr Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn 195 200 205

Val Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro 210 215 220

Arg Asp Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser 225 230 235 240

Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr 245 250 255

Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Ile Ile Lys Asp Asp 260 265 270

Pro Glu val Gin Phe Ser Trp Phe val Asp Asp Val Glu Val His Thr 275 280 285

Ala Gin Thr Gin Pro Arg Glu Gin Phe Asn Ser Thr Phe Arg Cheese 290 295 300

Val Ser Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Phe Lys Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys 325 330 335

Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys Al Pro Gin Val Tyr Thr 340 345 350

How much Pro Pro Pro Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr 355 360 365

Cys Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin

VP / 4572 / AGR

EP 2 569 010 B1

370

375

380

Trp Asn Gly Gin Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro Ile Met 385 390 395 400

Asp Thr Asp Gly Cheese Tyr Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys 405 410 415

Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu 420 425 430

Gly Leu His Asn His Thr Glu Lys Ser Leu Cheese His Ser Pro Gly 435 440 445

Lys <210> 35 <211> 11 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-D <400> 35

Gin Ala Ser Gin Gly Thr Ser Ile Asn Leu Asn 15 10 <210> 36 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-D <400> 36

Gly Cheese Ser Asn Leu Glu Asp

5 <210> 37 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-D <400> 37

Leu Gin His Cheese Tyr Leu Pro Tyr Thr 1 5 <210> 38 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-D <400> 38

Asp His Tyr Met Ser

5 <210> 39 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-D <400> 39

VP / 4572 / AGR

EP 2 569 010 B1

Asp Ile A3n Pro Tyr Ser Gly Glu Thr Thr Tyr Asn Gin Ly3 Phe Lys 15 10 15

Gly <210> 40 <211> 10 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-D <400> 40

Asp Asp Tyr Asp Ala Ser Pro Phe Ala Tyr

10 <210> 41 <211> 645 <212> DNA <213> Mus musculus <400> 41 gatgtccaga tgattcagtc tccatcctcc ctgtctgcat ctttgggaga catagtcacc 60 atgacttgcc aggcaagtca gggcactagc attaatttaa actggtttca gcaaaaacca 120 gggaaggctc ctaagctcct gatctatggt tcaagcaact tggaagatgg ggtcccatca 180 aggttcagtg gcagtagata tgggacagat ttcactctca ccatcagcag cctggaggat 240 gaagatctgg caacttattt ctgtctacaa catagttatc tcccgtacac gttcggaggg 300 gggaccaagc tggaaataaa acgggctgat gctgcaccaa ctgtatccat cttcccacca 360 tccagtgagc agttaacatc tggaggtgcc tcagtcgtgt gcttcttgaa caacttctac 420 cccaaagaca tcaatgtcaa gtggaagatt gatggcagtg aacgacaaaa tggcgtcctg 480 aacagttgga ctgatcagga cagcaaagac agcacctaca gcatgagcag caccctcacg 540 ttgaccaagg acgagtatga acgacataac agctatacct gtgaggccac tcacaagaca 600 tcaacttcac ccattgtcaa gagcttcaac aggaatgagt gttag 645 <210 >42 <211> 214 <212> PRT <213> Mus musculus <400> 42

VP / 4572 / AGR

EP 2 569 010 B1

Asp Val Gin Met Ile Gin Cheese Pro Cheese Cheese Leu Cheese Ala Cheese Leu Gly 15 10 15

Asp Ile Val Thr Met Thr Cys Gin Ala Ser Gin Gly Thr Ser Ile Asn 20 25 30

Leu Asn Trp Phe Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45

Tyr Gly Cheese Ser Asn Leu Glu Asp Gly Val Pro Cheese Arg Phe Cheese Gly 50 55 60

Ser Arg Tyr Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Asp 65 70 75 80

Glu Asp Leu Ala Thr Tyr Phe Cys Leu Gin His cheese Tyr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110

Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gin Leu Thr Cheese Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140

Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu 145 150 155 160

Asn Ser Trp Thr Asp Gin Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Cheese Tyr 180 185 190

Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205

Phe Asn Arg Asn Glu Cys 210 <210> 43 <211> 1332 <212> DNA <213> Mus musculus <400> 43

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggtccagc</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctggtgacgc</td><td>ctggggcttc</td><td>agtgaagata</td><td>60</td>

<td>tcttgtaagg</td><td>cttctggata</td><td>cacattcact</td><td>gaccactaca</td><td>tgagctgggt</td><td>gaagcagagt</td><td>120</td>

<td>catggaaaaa</td><td>gccttgagtg</td><td>gattggagat</td><td>attaatccct</td><td>attctggtga</td><td>aactacctac</td><td>180</td>

<td>aaccagaagt</td><td>tcaagggcac</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcttccag</td><td>tatagcctac</td><td>240</td>

<td>atggagatcc</td><td>gcggcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagagatgat</td><td>300</td>

<td>tacgacgcct</td><td>ctccgtttgc</td><td>ttactggggc</td><td>caagggactc</td><td>tggtcactgt</td><td>ctctgcagcc</td><td>360</td>

<td>aaaacgacac</td><td>ccccatctgt</td><td>ctatccactg</td><td>gcccctggat</td><td>ctgctgccca</td><td>aactaactcc</td><td>420</td>

<td>atggtgaccc</td><td>tgggatgcct</td><td>ggtcaagggc</td><td>tatttccctg</td><td>agccagtgac</td><td>agtgacctgg</td><td>480</td>

<td>aactctggat</td><td>ccctgtccag</td><td>cggtgtgcac</td><td>accttcccag</td><td>ctgtcctgca</td><td>gtctgacctc</td><td>540</td>

<td>tacactctga</td><td>gcagctcagt</td><td>gactgtcccc</td><td>tccagcacct</td><td>ggcccagcga</td><td>gaccgtcacc</td><td>600</td>

<td>tgcaacgttg</td><td>cccacccggc</td><td>cagcagcacc</td><td>aaggtggaca</td><td>agaaaattgt</td><td>gcccagggat</td><td>660</td>

<td>tgtggttgta</td><td>agccttgcat</td><td>atgtacagtc</td><td>ccagaagtat</td><td>catctgtctt</td><td>catcttcccc</td><td>720</td>

<td>ccaaagccca</td><td>aggatgtgct</td><td>caccattact</td><td>ctgactccta</td><td>aggtcacgtg</td><td>tgttgtggta</td><td>780</td>

<td>gacatcagca</td><td>aggatgatcc</td><td>cgaggtccag</td><td>ttcagctggt</td><td>ttgtagatga</td><td>tgtggaggtg</td><td>840</td>

<td>cacacagctc</td><td>agacgcaacc</td><td>ccgggaggag</td><td>cagttcaaca</td><td>gcactttccg</td><td>ctcagtcagt</td><td>900</td>

<td>gaacttccca</td><td>tcatgcacca</td><td>ggactggctc</td><td>aatggcaagg</td><td>agttcaaatg</td><td>cagggtcaac</td><td>960</td>

<td>agtccagctt</td><td>tccctgcccc</td><td>catcgagaaa</td><td>accatctcca</td><td>aaaccaaagg</td><td>cagaccgaag</td><td>1020</td>

<td>gctccacagg</td><td>tgtacaccat</td><td>tccacctccc</td><td>aaggagcaga</td><td>tggccaagga</td><td>taaagtcagt</td><td>1080</td>

<td>ctgacctgca</td><td>tgataacaga</td><td>cttcttccct</td><td>gaagacatta</td><td>ctgtggagtg</td><td>gcagtggaat</td><td>1140</td>

<td>gggcagccag</td><td>cggagaacta</td><td>caagaacact</td><td>cagcccatca</td><td>tggacacaga</td><td>tggctcttac</td><td>1200</td>

<td>ttcatctaca</td><td>gcaagctcaa</td><td>tgtgcagaag</td><td>agcaactggg</td><td>aggcaggaaa</td><td>tactttcacc</td><td>1260</td>

<td>tgctctgtgt</td><td>tacatgaggg</td><td>cctgcacaac</td><td>caccatactg</td><td>agaagagcct</td><td>ctcccactct</td><td>1320</td>

<td>cctggtaaat</td><td>ga</td><td></td><td></td><td></td><td></td><td>1332</td>

<210> 44 <211> 443 <212> PRT <213> Mus musculus <400> 44

VP / 4572 / AGR

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Cheese Gly Pro Glu Leu Val Thr Pro Gly Ala 15 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp His 20 25 30

Tyr Met Ser Trp Val Lys Gin Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45

Gly Asp Ile Asn Pro Tyr Cheese Gly Glu Thr Thr Tyr Asn Gin Ly3 Phe 50 55 60

Lys Gly Thr Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Ile Ala Tyr 65 70 75 80

Met Glu Ile Arg Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Asp Asp Tyr Asp Ala Ser Pro Phe Ala Tyr Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser Val Tyr 115 120 125

Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr Asn Cheese Met Val Thr Leu 130 135 140

Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp

VP / 4572 / AGR

EP 2 569 010 B1

145 150 155 160

Asn Cheese Gly Cheese Leu Cheese Cheese Gly Val His Thr Phe Pro Ala Val Leu 165 170 175

Gin Cheese Asp Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr Val Pro Ser Ser 180 185 190

Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala Hi3 Pro Ala Ser 195 200 205

Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys. Gly Cys Lys 210 215 220

Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro 225 230 235 240

Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr 245 250 255

Cys Val Val Val Asp. Ile Ser Lys. Asp Asp. Pro Glu. Val. Gin. Phe Ser 260 265 270

Trp Phe Val Asp Asp. Val. Glu. Val. His Thr. Ala Gin Thr. Gin Pro Arg. 275 280 285

Glu Glu Gin Phe Asn Cheese Thr Phe Arg Cheese Val Ser Glu Leu Pro Ile 290 295 300

Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Lys Cy3 Arg Val Asn 305 310 315 320

Pro Pro Ala Pro Phe Pro Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys 325 330 335

Gly Arg Pro Lys Ala Pro Gin Val Tyr Thr How much Pro Pro Pro Lys Glu 340 345 350

Gin Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe 355 360 365

Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn Gly Gin Pro Ala 370 375 380

Glu Asn Tyr Lys Asn Thr Gin Pro How Much Met Asp Thr Asp Gly Cheese Tyr

VP / 4572 / AGR

EP 2 569 010 B1

385 390 395 400

Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys Cheese Asn Trp Glu Ala Gly 405 410 415

Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His 420 425 430

Thr Glu Lys Ser Leu Cheese His Ser Pro Gly Lys 435 440 <210> 45 <211> 11 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-2 <400> 45

Arg Ala Ser Ser Ser Val Tyr Tyr Tyr Met His

10 <210> 46 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-2 <400> 46

Ala Thr Ser Asn Leu Ala Ser 1 5 <210> 47 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-3 <400> 47

Gin Gin Trp Cheese Ser Asp Pro Leu Thr 1 5 <210> 48 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-2 <400> 48

Asp Tyr Phe Ile His

5 <210> 49 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-2 <400> 49

Arg Leu Asp Pro Glu Asp Gly Glu Asp Asp Tyr Al Pro Pro Lys Phe Gin 15 10 15

Asp <210> 50 <211> 12

VP / 4572 / AGR

EP 2 569 010 B1 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-2 <400> 50

Glu Asp Tyr Asp Gly Thr Tyr Thr Phe Phe Pro Tyr

10 <210> 51 <211> 642 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> light chain of the Ab-2 (no signal) <400> 51 caaattgttc tctcccagtc tccagcaatc ctgtctacat ctccagggga gaaggtcaca 60 atgacttgca gggccagctc aagtgtatat tacatgcact ggtaccagca gaagccagga 120 tcctccccca aaccctggat ttatgccaca tccaacctgg cttctggagt ccctgttcgc 180 ttcagtggca gtgggtctgg gacctcttac tctctcacaa tcaccagagt ggaggctgaa 240 gatgctgcca cttattactg ccagcagtgg agtagtgacc cactcacgtt cggtgctggg 300 accaagctgg agctgaaacg ggctgatgct gcaccaactg tatccatctt cccaccatcc 360 agtgagcagt taacatctgg aggtgcctca gtcgtgtgct tcttgaacaa cttctacccc 420 aaagacatca atgtcaagtg gaagattgat ggcagtgaac gacaaaatgg cgtcctgaac 480 agttggactg atcaggacag caaagacagc acctacagca tgagcagcac cctcacgttg 540 accaaggacg agtatgaacg acataacagc tatacctgtg aggccactca caagacatca 600 acttcaccca ttgtcaagag cttcaacagg aatgagtgtt ag 642 <210> 52 <211> 213 <212>PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-2 light chain (no signal) <400> 52

VP / 4572 / AGR

EP 2 569 010 B1

Gin Ile Val Leu Cheese Gin Cheese Pro Ala Ile Leu Cheese Thr Cheese Pro Gly 15 10 15

Glu Lys Val Thr Met Thr Cys Arg Ala Cheese Cheese Cheese Val Tyr Tyr Met 20 25 30

His Trp Tyr Gin Gin Ly3 Pro Gly Cheese Pro Pro Lys Pro Trp Ile Tire 35 40 45

Ala Thr Cheese Asn Leu Ala Cheese Gly Val Pro Val Arg Phe Cheese Gly Cheese 50 55 60

Gly Cheese Gly Thr Cheese Tyr Cheese Leu Thr Ile Thr Arg Val Glu Ala Glu 65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cy3 Gin Gin Trp Cheese Cheese Asp Pro Leu Thr 85 90 95

Phe Gly Ala Gly Thr Ly3 Leu Glu Leu Lys Arg Ala Asp Ala Ala Pro 100 105 110

Thr Val Ser Ile Phe Pro Pro Cheese Ser Glu Gin Leu Thr Cheese Gly Gly 115 120 125

Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp. Asn 130 135 140

Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu Asn 145 150 155 160

Trp cheese Asp Asp. Asp cheese Asp. Lys Asp Ser Thr Tyr Cheese Met Ser Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr Thr 180 185 190

Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser Phe 195 200 205

Asn Arg Asn Glu Cys 210 <210> 53 <211> 1338 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-2 heavy chain (no signal) <400> 53

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggttcagg</td><td>tgcagcagtc</td><td>tgggccagaa</td><td>cttgtgaagc</td><td>caggggcctc</td><td>agtcaagttg</td><td>60</td>

<td>tcctgcacag</td><td>cttctggctt</td><td>caacattaaa</td><td>gactacttta</td><td>tacactgggt</td><td>gaagcagagg</td><td>120</td>

<td>cctgaacagg</td><td>gcctggagtg</td><td>gattggaagg</td><td>cttgatcctg</td><td>aggatggtga</td><td>aagtgattat</td><td>180</td>

<td>gccccgaagt</td><td>tccaggacaa</td><td>ggccattatg</td><td>acagcagaca</td><td>catcatccaa</td><td>cacagcctat</td><td>240</td>

<td>cttcagctca</td><td>gaagcctgac</td><td>atctgaggac</td><td>actgccatct</td><td>attattgtga</td><td>gagagaggac</td><td>300</td>

<td>tacgatggta</td><td>cctacacctt</td><td>ttttccttac</td><td>tggggccaag</td><td>ggactctggt</td><td>cactgtctct</td><td>360</td>

<td>gcagccaaaa</td><td>cgacaccccc</td><td>atctgtctat</td><td>ccactggccc</td><td>ctggatctgc</td><td>tgcccaaact</td><td>420</td>

<td>aactccatgg</td><td>tgaccctggg</td><td>atgcctggtc</td><td>aagggctatt</td><td>tccctgagcc</td><td>agtgacagtg</td><td>480</td>

<td>acctggaact</td><td>ctggatccct</td><td>gtccagcggt</td><td>gtgcacacct</td><td>tcccagctgt</td><td>cctgcagtct</td><td>540</td>

<td>gacctctaca</td><td>ctctgagcag</td><td>ctcagtgact</td><td>gtcccctcca</td><td>gcacctggcc</td><td>cagcgagacc</td><td>600</td>

<td>gtcacctgca</td><td>acgttgccca</td><td>cccggccagc</td><td>agcaccaagg</td><td>tggacaagaa</td><td>aattgtgccc</td><td>660</td>

<td>agggattgtg</td><td>gttgtaagcc</td><td>ttgcatatgt</td><td>acagtcccag</td><td>aagtatcatc</td><td>tgtcttcatc</td><td>720</td>

<td>ttccccccaa</td><td>agcccaagga</td><td>tgtgctcacc</td><td>attactctga</td><td>ctcctaaggt</td><td>cacgtgtgtt</td><td>780</td>

<td>gtggtagaca</td><td>tcagcaagga</td><td>tgatcccgag</td><td>gtccagttca</td><td>gctggtttgt</td><td>agatgatgtg</td><td>840</td>

<td>gaggtgcaca</td><td>cagctcagac</td><td>gcaaccccgg</td><td>gaggagcagt</td><td>tcaacagcac</td><td>tttccgctca</td><td>900</td>

<td>gtcagtgaac</td><td>ttcccatcat</td><td>gcaccaggac</td><td>tggctcaatg</td><td>gcaaggagtt</td><td>caaatgcagg</td><td>960</td>

<td>gtcaacagtg</td><td>cagctttccc</td><td>tgcccccatc</td><td>gagaaaacca</td><td>tctccaaaac</td><td>caaaggcaga</td><td>1020</td>

<td>ccgaaggctc</td><td>cacaggtgta</td><td>caccattcca</td><td>cctcccaagg</td><td>agcagatggc</td><td>caaggataaa</td><td>1080</td>

<td>gtcagtctga</td><td>cctgcatgat</td><td>aacagacttc</td><td>ttccctgaag</td><td>acattactgt</td><td>ggagtggcag</td><td>1140</td>

<td>tggaatgggc</td><td>agccagcgga</td><td>gaactacaag</td><td>aacactcagc</td><td>ccatcatgga</td><td>cacagatggc</td><td>1200</td>

<td>tcttacttca</td><td>tctacagcaa</td><td>gctcaatgtg</td><td>cagaagagca</td><td>actgggaggc</td><td>aggaaatact</td><td>1260</td>

<td>ttcacctgct</td><td>ctgtgttaca</td><td>tgagggcctg</td><td>cacaaccacc</td><td>atactgagaa</td><td>gagcctctcc</td><td>1320</td>

<td>cactctcctg</td><td>gtaaatga</td><td></td><td></td><td></td><td></td><td>1338</td>

<210> 54 <211> 445 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-2 heavy chain (no signal) <400> 54

VP / 4572 / AGR

EP 2 569 010 B1

Glu Val Gin Val Gin Gin Gly Pro Ser Glu Leu Val Lys Pro Gly Ala 15 10 15

Cheese Val Lys Leu Cheese Cys Thr Ala Cheese Gly Phe Asn Ile Lys Asp Tyr 20 25 30

Phe Ile His Trp Val Lys Gin Arg Pro Glu Gin Gly Leu Glu Trp Ile 35 40 45

Gly Arg Leu Asp Pro Glu Asp Gly Glu Asp Asp Tyr Ala Pro Lys Phe 50 55 60

Gin Asp Lys Ala Ile Met Thr Ala Asp Thr Cheese Ser Asn Thr Ala Tyr 65 70 75 80

Leu Gin Leu Arg Leu Ser Thr Cheese Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95

Glu Arg Glu Asp Tyr Asp Gly Thr Tyr Thr Phe Phe Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser

VP / 4572 / AGR

EP 2 569 010 B1

115 120 125

Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr A3n Ser Met Val 130 135 140

Thr Leu Gly Cy3 Leu Val Ly3 Gly Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Thr Trp Asn Cheese Gly Cheese Leu Cheese Cheese Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Cheese Asp Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr Val Pro 180 185 190

Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cy3 Asn Val Ala His Pro 195 200 205

Ala Ser Ser Thr Lys Val Asp Lys Ly3 Ile Val Pro Arg Asp. Cys Gly 210 215 220

Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile 225 230 235 240

Phe Pro Pro Ly3 Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys 245 250 255

Val Thr Cys Val Val Val Asp. Ile Ser Lys Asp Asp Pro. Glu Val. Gin 260 265 270

Phe Ser Trp Phe Val Asp Asp. Val. Glu. Val. His Thr Ala Gin Thr. Gin 275 280 285

Pro Arg Glu Glu Gin Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu 290 295 300

Pro Ile Met His Gin Asp Trp Leu A3n Gly Lys Glu Phe Lys Cys Arg 305 310 315 320

Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Ile Glu Lys Thr Ile Ser Lys 325 330 335

Thr Lys Gly Arg Pro Lys Ala Pro Gin Val Tyr Thr How much Pro Pro Pro 340 345 350

Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr

VP / 4572 / AGR

EP 2 569 010 B1

355

360

365

Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn Gly Gin 370 375 380

Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro How Much Met Asp Thr Asp Gly 385 390 395 400

Cheese Tyr Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys Cheese Asn Trp Glu 405 410 415

Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn 420 425 430

His His Thr Glu Lys Ser Leu Ser His Cheese Pro Gly Lys 435 440 445 <210> 55 <211> 12 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-3 and Ab-15 <400> 55

Ser Val Ser Ser Thr Ile Ser Ser Asn His Leu Hi3 15 10 <210> 56 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-3 and Ab-15 <400> 56

Gly Thr Ser Asn Leu Ala Ser

5 <210> 57 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-3 and Ab-15 <400> 57

Gin Gin Trp Cheese Cheese Tyr Pro Leu Thr

5 <210> 58 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-3 and Ab-15 <400> 58

Asp Phe Tyr Leu Hi3 1 5 <210> 59 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-3 and Ab-15 <400> 59

VP / 4572 / AGR

EP 2 569 010 B1

Arg Ile Asp Pro Glu Asn Gly Asp Thr Leu Tyr Asp Pro Lys Phe Gin 15 10 15

Asp <210> 60 <211> 16 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-3 and Ab-15 <400> 60

Glu Ala Asp Tyr Phe His Asp Gly Thr Ser Tyr Trp Tyr Phe Asp Val 15 10 15 <210> 61 <211> 648 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-3 light chain <400> 61

<td>gaaattgtgc</td><td>tcacccagtc</td><td>tccagcactc</td><td>atggctgcat</td><td>ctccggggga</td><td>gaaggtcacc</td><td>60</td>

<td>atcacctgca</td><td>gtgtcagttc</td><td>aactataagt</td><td>tccaaccact</td><td>tgcactggtt</td><td>ccagcagaag</td><td>120</td>

<td>tcagacacct</td><td>cccccaaacc</td><td>ctggatttat</td><td>ggcacatcca</td><td>acctggcttc</td><td>tggagtccct</td><td>180</td>

<td>gttcgcttca</td><td>gtggcagtgg</td><td>atctgggacc</td><td>tcttattctc</td><td>tcacaatcag</td><td>cagcatggag</td><td>240</td>

<td>gctgaggatg</td><td>ctgccactta</td><td>ttactgtcaa</td><td>cagtggagta</td><td>gttacccact</td><td>cacgttcggc</td><td>300</td>

<td>gctgggacca</td><td>agctggagct</td><td>gagacgggct</td><td>gatgctgcac</td><td>caactgtatc</td><td>catcttccca</td><td>360</td>

<td>ccatccagtg</td><td>agcagttaac</td><td>atctggaggt</td><td>gcctcagtcg</td><td>tgtgcttctt</td><td>gaacaacttc</td><td>420</td>

<td>taccccaaag</td><td>acatcaatgt</td><td>caagtggaag</td><td>attgatggca</td><td>gtgaacgaca</td><td>aaatggcgtc</td><td>480</td>

<td>ctgaacagtt</td><td>ggactgatca</td><td>ggacagcaaa</td><td>gacagcacct</td><td>acagcatgag</td><td>cagcaccctc</td><td>540</td>

<td>acgttgacca</td><td>aggacgagta</td><td>tgaacgacat</td><td>aacagctata</td><td>cctgtgaggc</td><td>cactcacaag</td><td>600</td>

<td>acatcaactt</td><td>cacccattgt</td><td>caagagcttc</td><td>aacaggaatg</td><td>agtgttag</td><td></td><td>648</td>

<210> 62 <211> 215 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Light chain Ab-3 (no signal) <400> 62

VP / 4572 / AGR

EP 2 569 010 B1

Glu Ile Val Leu Thr Gin 1 5

Pro Pro Ala Leu Met 10

Ala Ala Ser Pro Gly 15

Glu Lys Val Thr Ile Thr 20

Cys Ser Val Ser Ser 25

Thr Ile Ser Ser Asn 30

His Leu His Trp Phe Gin 35

Gin Lys Ser A3p Thr 40

Pro Pro Ly3 Pro Trp 45 cheese

How Many Tyr Gly Thr Ser Asn 50

Leu Ala Ser Gly Val 55

Pro Val Arg Phe Ser 60

Gly Cheese Gly Cheese Gly Thr 65 70

Cheese Tyr Ser Leu Thr 75

Ile Ser Ser Met Glu 80

Ala Glu Asp Ala Ala Thr 85

Tyr Tyr Cys Gin Gin 90

Trp Cheese Cheese Tyr Pro 95

Leu Thr Phe Gly Ala Gly 100

Thr Lys Leu Glu Leu 105

Arg Arg Ala Asp Ala 110

Ala Pro Thr Val Ser Ile 115

Phe Pro Pro Cheese Cheese 120

Glu Gin Leu Thr Ser 125

Gly Gly Ala Ser Val Val 130

Cys Phe Leu Asn Asn 135

Phe Tyr Pro Lys Asp 140

Ile Asn Val Lys Trp Lys 145 150

Ile Asp Gly Ser Glu 155

Arg Gin Asn Gly Val 160

Leu Asn Cheese Trp Thr Asp 165

Gin Asp Ser Lys Asp 170

Cheese Thr Tyr Cheese Met 175

Ser Ser Thr Leu Thr Leu 180

Thr Lys Asp Glu Tyr 185

Glu Arg His Asn Cheese 190

Tyr Thr Cys Glu Ala Thr 195

His Lys Thr Ser Thr 200

Pro Pro Ile Val Lys 205

Ser Phe Asn Arg Asn Glu 210 <210> 63 <211> 1350 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-3 heavy chain <400> 63

Cys

215

VP / 4572 / AGR

EP 2 569 010 B1 gaggttcagc tgcagcagtc tggggctgaa cttgtgaggc caggggcctt agtcaagttg 60 tcctgcacag cttctgactt caacattaaa gacttctatc tacactggat gaggcagcgg 120 cctgaacagg gcctggactg gattggaagg attgatcctg agaatggtga tactttatat 180 gacccgaagt tccaggacaa ggccactctt acaacagaca catcctccaa cacagcctac 240 ctgcagctca gcggcctgac atctgagacc actgccgtct attactgttc tagagaggcg 300 gattatttcc acgatggtac ctcctactgg tacttcgatg tctggggcgc agggaccaca 360 atcaccgtct cctcagccaa aacgacaccc ccatctgtct atccactggc ccctggatct 420 gctgcccaaa ctaactccat ggtgaccctg ggatgcctgg tcaagggcta tttccctgag 480 ccagtgacag tgacctggaa ctctggatcc ctgtccagcg gtgtgcacac cttcccagct 540 gtcctgcagt ctgacctcta cactctgagc agctcagtga ctgtcccctc cagcacctgg 600 cccagcgaga ccgtcacctg caacgttgcc cacccggcca gcagcaccaa ggtggacaag 660 aaaattgtgc ccagggattg tggttgtaag ccttgcatat gtacagtccc agaagtatca 720tctgtcttca tcttcccccc aaagcccaag gatgtgctca ccattactct gactcctaag 780 gtcacgtgtg ttgtggtaga catcagcaag gatgatcccg aggtccagtt cagctggttt 840 gtagatgatg tggaggtgca cacagctcag acgcaacccc gggaggagca gttcaacagc 900 actttccgct cagtcagtga acttcccatc atgcaccagg actggctcaa tggcaaggag 960 ttcaaatgca gggtcaacag tgcagctttc cctgccccca tcgagaaaac catctccaaa 1020 accaaaggca gaccgaaggc tccacaggtg tacaccattc cacctcccaa ggagcagatg 1080 gccaaggata aagtcagtct gacctgcatg ataacagact tcttccctga agacattact 1140 gtggagtggc agtggaatgg gcagccagcg gagaactaca agaacactca gcccatcatg 1200 gacacagatg gctcttactt catctacagc aagctcaatg tgcagaagag caactgggag 1260 gcaggaaata ctttcacctg ctctgtgtta catgagggcc tgcacaacca ccatactgag 1320 aagagcctct cccactctcc tggtaaatga 1350 <210> 64 <211> 449 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-3 heavy chain (no signal) <400> 64

VP / 4572 / AGR

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Cheese Gly Ala Glu Leu Val Arg Pro Gly Ala 15 10 15

Leu Val Lys Leu Cheese Cys Thr Ala Cheese Asp Phe Asn Ile Lys Asp Phe 20 25 30

Tyr Leu His Trp Met Arg Arg Arg Arg Gin Gly Leu Asp Trp Ile 35 40 45

Gly Arg Ile Ile Pro Glu Asn Gly Asp Thr Leu Tyr Asp Pro Lys Phe 50 55 60

Gin Asp Lys Ala Thr Leu Thr Thr Asp Thr Cheese Ser Asn Thr Ala Tyr 65 70 75 80

Leu Gin Leu Cheese Gly Leu Thr Cheese Glu Thr Thr Ala Val Tyr Tyr Cys

VP / 4572 / AGR

EP 2 569 010 B1

90 95

Ser Arg Glu Ala Asp Tyr Phe His Asp Gly Thr Ser Tyr Trp Tyr Phe 100 105 110

Asp Val Trp Gly Ala Gly Thr Thr Ile Thr Val Ser Ser Ala Lys Thr 115 120 125

Thr Pro Pro Ser Val Tire Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr 130 135 140

Asn Cheese Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu 145 150 155 160

Pro Val Thr Val Thr Trp A3n Ser Gly Ser Leu Ser Ser Gly Val His 165 Π0 175

Thr Phe Pro Ala Val Leu Gin Le Asp cheese Leu Tyr Thr Leu Ser Ser Ser 180 185 190

Val Thr Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn 195 200 205

Val Ala Hi3 Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro 210 215 220

Arg Asp Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser 225 230 235 240

Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr 245 250 255

Leu Thr Pro Lye Val Thr Cys Val Val Val A3p Ile Ser Ly3 Asp Asp 260 265 270

Pro Glu Val Gin Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr 275 280 285

Ala Gin Thr Gin Pro Arg Glu Gin Phe Asn Ser Thr Phe Arg Cheese 290 295 300

Val Ser Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu 305 310 315 320

Phe Lys Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys

VP / 4572 / AGR

EP 2 569 010 B1

325 330 335

Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys Al Pro Gin Val Tyr Thr 340 345 350

How much Pro Pro Pro Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr 355 360 365

Cys Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin 370 375 380

Trp Asn Gly Gin Pro Ala Glu Asn Tyr Lys A3n Thr Gin Pro Ile Met 385 390 395 400

Asp Thr Asp Gly Cheese Tyr Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys 405 410 415

Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu 420 425 430

Gly Leu His Asn His Thr Glu Lys Ser Leu Cheese His Ser Pro Gly 435 440 445

Lys <210> 65 <211> 324 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Lightweight variable Ab-15 <400> 65

<td>gacatccaga</td><td>tgacccagtc</td><td>tccatcctcc</td><td>ctctcagcat</td><td>ccgtaggcga</td><td>tagagttaca</td><td>60</td>

<td>ataacatgca</td><td>gcgtatcatc</td><td>aactatatca</td><td>tcaaatcatc</td><td>ttcattggtt</td><td>ccaacagaaa</td><td>120</td>

<td>cccggcaaag</td><td>cacctaaatc</td><td>acttatatac</td><td>ggcacatcaa</td><td>atctcgcatc</td><td>aggcgttcct</td><td>180</td>

<td>tcaagatttt</td><td>caggctctgg</td><td>ctcaggcacc</td><td>gactttactc</td><td>ttacaatatc</td><td>ctccctccaa</td><td>240</td>

<td>cccgaagact</td><td>tcgcaaccta</td><td>ttactgtcaa</td><td>caatggtcct</td><td>catatccact</td><td>cacatttggc</td><td>300</td>

<td>ggcggcacaa</td><td>aagtagaaat</td><td>yeah</td><td></td><td></td><td></td><td>324</td>

<210> 66 <211> 108 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN. <223> Lightweight variable Ab-15 <400> 66

VP / 4572 / AGR

EP 2 569 010 B1

Asp Ile Gin Met Thr Gin 1 5

Cheese Pro Cheese Cheese Leu Cheese Ala Cheese Val Gly 10 15

Asp Arg Val Thr Ile Thr 20

Cys Ser Val Ser Ser Thr Ile Ser Ser Asn 25 30

His Leu His Trp Phe Gin 35

Gin Lys Pro Gly Lys Ala Pro Lys Ser Leu 40 45

How Many Tyr Gly Thr Ser Asn 50

Leu Ala Ser Gly Val Pro Ser Arg Phe Cheese 55 60

Gly Cheese Gly Cheese Gly Thr 65 70

Asp Phe Thr Leu Thr Ile Ser Ser Leu Gin 75 80

Pro Glu A3p Phe Ala Thr 85

Tyr Tyr Cys Gin Gin Trp. Cheese Ser Tyr Pro 90 95

Leu Thr Phe Gly Gly Gly 100 <210> 67 <211> 375 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Heavy variable Ab-15

Thr Ly3 Val Glu Ile Lys 105 <400> 67

<td>gaggtgcagc</td><td>tggtgcagtc</td><td>tggggctgag</td><td>gtgaagaagc</td><td>ctggggcctc</td><td>agtgaaggtc</td><td>60</td>

<td>tcctgcaagg</td><td>cttctgactt</td><td>caacattaaa</td><td>gacttctatc</td><td>tacactgggt</td><td>gcgacaggcc</td><td>120</td>

<td>cctggacaag</td><td>ggcttgagtg</td><td>gattggaagg</td><td>attgatcctg</td><td>agaatggtga</td><td>tactttatat</td><td>180</td>

<td>gacccgaagt</td><td>tccaggacaa</td><td>ggtcaccatg</td><td>accacagaca</td><td>cgtccaccag</td><td>cacagcctac</td><td>240</td>

<td>atggagctga</td><td>ggagcctgag</td><td>atctgacgac</td><td>acggccgtgt</td><td>attactgtgc</td><td>gagagaggcg</td><td>300</td>

<td>gattatttcc</td><td>acgatggtac</td><td>ctcctactgg</td><td>tacttcgatg</td><td>tctggggccg</td><td>tggcaccctg</td><td>360</td>

gtcaccgtct ctagt 375 <210> 68 <211> 125 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN. <223> Heavy variable Ab-15 <400> 68

VP / 4572 / AGR

EP 2 569 010 B1

Glu Val Gin Leu Val Gin Cheese Gly Ala Glu Val Lys Pro Gly Ala 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Asp Phe Asn Ile Lye Asp Phe 20 25 30

Tyr Leu His Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Ile 35 40 45

Gly Arg Ile Ile Pro Glu Asn Gly Asp Thr Leu Tyr Asp Pro Lys Phe 50 55 60

Gin Asp Lys Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Leu Arg Ser Asp Asp Thr Al Val Tyr Tyr Cys 85 90 95

Ala Arg Glu Ala Asp Tyr Phe His Asp Gly Thr Ser Tyr Trp Tyr Phe 100 105 110

Asp Val Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 69 <211> 645 <212> DNA <213> Artificial Sequence <220>

<223> Humanized antibody <220>

<221> trait_different

<223> Ab-15 light chain <400> 69

<td>gacatccaga</td><td>tgacccagtc</td><td>tccatcctcc</td><td>ctctcagcat</td><td>ccgtaggcga</td><td>tagagttaca</td><td></td>

<td>ataacatgca</td><td>gcgtatcatc</td><td>aactatatca</td><td>tcaaatcatc</td><td>ttcattggtt</td><td>ccaacagaaa</td><td>120</td>

<td>cccggcaaag</td><td>cacctaaatc</td><td>acttatatac</td><td>ggcacatcaa</td><td>atctcgcatc</td><td>aggcgttcct</td><td>180</td>

<td>tcaagatttt</td><td>caggctctgg</td><td>ctcaggcacc</td><td>gactttactc</td><td>ttacaatatc</td><td>ctccctccaa</td><td>240</td>

<td>cccgaagact</td><td>tcgcaaccta</td><td>ttactgtcaa</td><td>caatggtcct</td><td>catatccact</td><td>cacatttggc</td><td>300</td>

<td>ggcggcacaa</td><td>aagtagaaat</td><td>taaacgtacg</td><td>gtggctgcac</td><td>catctgtctt</td><td>catcttcccg</td><td>360</td>

<td>ccatctgatg</td><td>agcagttgaa</td><td>atctggaact</td><td>gcctctgttg</td><td>tgtgcctgct</td><td>gaataacttc</td><td>420</td>

<td>tatcccagag</td><td>aggccaaagt</td><td>acagtggaag</td><td>gtggataacg</td><td>ccctccaatc</td><td>gggtaactcc</td><td>480</td>

<td>caggagagtg</td><td>tcacagagca</td><td>ggacagcaag</td><td>gacagcacct</td><td>acagcctcag</td><td>cagcaccctg</td><td>540</td>

<td>acgctgagca</td><td>aagcagacta</td><td>cgagaaacac</td><td>aaagtctacg</td><td>cctgcgaagt</td><td>cacccatcag</td><td>600</td>

<td>ggcctgagct</td><td>cgcccgtcac</td><td>aaagagcttc</td><td>aacaggggag</td><td>agtgt</td><td></td><td>645</td>

<210> 70 <211> 215 <212> PRT <213> Artificial sequence

VP / 4572 / AGR

EP 2 569 010 B1 <220>

<223> Sequence of humanized antibody <220>

<221> CECHA_RÓŻN.

<223> Ab-15 light chain

<td colspan="2">&lt;400&gt; 70</td><td colspan="2" rowspan="2">Gin Met</td><td rowspan="2">Thr 5</td><td rowspan="2">Gin</td><td rowspan="2">Cheese</td><td rowspan="2">Pro</td><td rowspan="2">Cheese</td><td rowspan="2">Cheese 10</td><td rowspan="2">Leu</td><td rowspan="2">Cheese</td><td rowspan="2">ala</td><td rowspan="2">Cheese</td><td rowspan="2">val 15</td><td rowspan="2">Gly</td>

<td>Asp 1</td><td>How much</td>

<td>Asp</td><td>Arg</td><td>val</td><td>Thr</td><td>How much</td><td>Thr</td><td>Cy3</td><td>Cheese</td><td>val</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>own</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>His</td><td>Leu</td><td>His</td><td>Trp</td><td>phe</td><td>Gin</td><td>Gin</td><td>Ly3</td><td>Pro</td><td>Gly</td><td>Lye</td><td>ala</td><td>Pro</td><td>lys</td><td>Cheese</td><td>Leu</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>How much</td><td>Tyr</td><td>Gly</td><td>Thr</td><td>Cheese</td><td>own</td><td>Leu</td><td>ala</td><td>Cheese</td><td>Gly</td><td>val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>phe</td><td>Cheese</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Gly</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Pro</td><td>Glu</td><td>Asp</td><td>phe</td><td>ala</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Trp</td><td>Cheese</td><td>Cheese</td><td>Tyr</td><td>Pro</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>Leu</td><td>Thr</td><td>phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>lys</td><td>val</td><td>Glu</td><td>How much</td><td>lys</td><td>Arg</td><td>Thr</td><td>val</td><td>ala</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>ala</td><td>Pro</td><td>Cheese</td><td>val</td><td>phe</td><td>How much</td><td>phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>Glu</td><td>Gin</td><td>Leu</td><td>lys</td><td>Cheese</td>

<td></td><td></td><td>115</td><td></td><td></td><td></td><td></td><td>120</td><td></td><td></td><td></td><td></td><td>125</td><td></td><td></td><td></td>

<td>Gly</td><td>Thr</td><td>ala</td><td>Cheese</td><td>val</td><td>val</td><td>Cys</td><td>Leu</td><td>Leu</td><td>own</td><td>own</td><td>phe</td><td>Tyr</td><td>Pro</td><td>Arg</td><td>Glu</td>

<td></td><td>130</td><td></td><td></td><td></td><td></td><td>135</td><td></td><td></td><td></td><td></td><td>140</td><td></td><td></td><td></td><td></td>

<td>ala</td><td>lys</td><td>val</td><td>Gin</td><td>Trp</td><td>lys</td><td>val</td><td>Asp</td><td>own</td><td>ala</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Gly</td><td>own</td><td>Cheese</td>

<td>145</td><td></td><td></td><td></td><td></td><td>150</td><td></td><td></td><td></td><td></td><td>155</td><td></td><td></td><td></td><td></td><td>160</td>

<td>Gin</td><td>Glu</td><td>Cheese</td><td>val</td><td>Thr</td><td>Glu</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>lys</td><td>Asp</td><td>Cheese</td><td>Thr</td><td>Tyr</td><td>Cheese</td><td>Leu</td>

<td></td><td></td><td></td><td></td><td>165</td><td></td><td></td><td></td><td></td><td>170</td><td></td><td></td><td></td><td></td><td>175</td><td></td>

<td>Cheese</td><td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>lys</td><td>ala</td><td>Asp</td><td>Tyr</td><td>Glu</td><td>lys</td><td>His</td><td>lys</td><td>val</td>

<td></td><td></td><td></td><td>180</td><td></td><td></td><td></td><td></td><td>185</td><td></td><td></td><td></td><td></td><td>190</td><td></td><td></td>

<td>Tyr</td><td>ala</td><td>Cys</td><td>Glu</td><td>val</td><td>Thr</td><td>His</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Pro</td><td>val</td><td>Thr</td><td>lys</td>

<td></td><td></td><td>195</td><td></td><td></td><td></td><td></td><td>200</td><td></td><td></td><td></td><td></td><td>205</td><td></td><td></td><td></td>

<td>Cheese</td><td>phe</td><td>own</td><td>Arg</td><td>Gly</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>210</td><td></td><td></td><td></td><td></td><td>215</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<210> 71 <211> 1353 <212> DNA <213> Artificial sequence <220>

VP / 4572 / AGR

EP 2 569 010 B1 <223> Humanized antibody <220>

<221> trait_different

<223> Ab-15 heavy chain <400> 71

<td>gaggtgcagc</td><td>tggtgcagtc</td><td>tggggctgag</td><td>gtgaagaagc</td><td>ctggggcctc</td><td>agtgaaggtc</td><td>60</td>

<td>tcctgcaagg</td><td>cttctgactt</td><td>caacattaaa</td><td>gacttctatc</td><td>tacactgggt</td><td>gcgacaggcc</td><td>120</td>

<td>cctggacaag</td><td>ggcttgagtg</td><td>gattggaagg</td><td>attgatcctg</td><td>agaatggtga</td><td>tactttatat</td><td>180</td>

<td>gacccgaagt</td><td>tccaggacaa</td><td>ggtcaccatg</td><td>accacagaca</td><td>cgtccaccag</td><td>cacagcctac</td><td>240</td>

<td>atggagctga</td><td>ggagcctgag</td><td>atctgacgac</td><td>acggccgtgt</td><td>attactgtgc</td><td>gagagaggcg</td><td>300</td>

<td>gattatttcc</td><td>acgatggtac</td><td>ctcctactgg</td><td>tacttcgatg</td><td>tctggggccg</td><td>tggcaccctg</td><td>360</td>

<td>gtcaccgtct</td><td>ctagtgcctc</td><td>caccaagggc</td><td>ccatcggtct</td><td>tccccctggc</td><td>gccctgctcc</td><td>420</td>

<td>aggagcacct</td><td>ccgagagcac</td><td>agcggccctg</td><td>ggctgcctgg</td><td>tcaaggacta</td><td>cttccccgaa</td><td>480</td>

<td>ccggtgacgg</td><td>tgtcgtggaa</td><td>ctcaggcgct</td><td>ctgaccagcg</td><td>gcgtgcacac</td><td>cttcccagct</td><td>540</td>

<td>gtcctacagt</td><td>cctcaggact</td><td>ctactccctc</td><td>agcagcgtgg</td><td>tgaccgtgcc</td><td>ctccagcaac</td><td>600</td>

<td>ttcggcaccc</td><td>agacctacac</td><td>ctgcaacgta</td><td>gatcacaagc</td><td>ccagcaacac</td><td>caaggtggac</td><td>660</td>

<td>aagacagttg</td><td>agcgcaaatg</td><td>ttgtgtcgag</td><td>tgcccaccgt</td><td>gcccagcacc</td><td>acctgtggca</td><td>720</td>

<td>ggaccgtcag</td><td>tcttcctctt</td><td>ccccccaaaa</td><td>cccaaggaca</td><td>ccctcatgat</td><td>ctcccggacc</td><td>780</td>

<td>cctgaggtca</td><td>cgtgcgtggt</td><td>ggtggacgtg</td><td>agccacgaag</td><td>accccgaggt</td><td>ccagttcaac</td><td>840</td>

<td>tggtacgtgg</td><td>acggcgtgga</td><td>ggtgcataat</td><td>gccaagacaa</td><td>agccacggga</td><td>ggagcagttc</td><td>900</td>

<td>aacagcacgt</td><td>tccgtgtggt</td><td>cagcgtcctc</td><td>accgttgtgc</td><td>accaggactg</td><td>gctgaacggc</td><td>960</td>

<td>aaggagtaca</td><td>agtgcaaggt</td><td>ctccaacaaa</td><td>ggcctcccag</td><td>cccccatcga</td><td>gaaaaccatc</td><td>1020</td>

<td>tccaaaacca</td><td>aagggcagcc</td><td>ccgagaacca</td><td>caggtgtaca</td><td>ccctgccccc</td><td>atcccgggag</td><td>1080</td>

<td>gagatgacca</td><td>agaaccaggt</td><td>cagcctgacc</td><td>tgcctggtca</td><td>aaggcttcta</td><td>ccccagcgac</td><td>1140</td>

<td>atcgccgtgg</td><td>agtgggagag</td><td>caatgggcag</td><td>ccggagaaca</td><td>actacaagac</td><td>cacacctccc</td><td>1200</td>

<td>atgctggact</td><td>ccgacggctc</td><td>cttcttcctc</td><td>tacagcaagc</td><td>tcaccgtgga</td><td>caagagcagg</td><td>1260</td>

<td>tggcagcagg</td><td>ggaacgtctt</td><td>ctcatgctcc</td><td>gtgatgcatg</td><td>aggctctgca</td><td>caaccactac</td><td>1320</td>

<td>acgcagaaga</td><td>gcctctccct</td><td>gtctccgggt</td><td>aaa</td><td></td><td></td><td>1353</td>

<210> 72 <211> 451 <212> PRT <213> Artificial sequence <220>

<223> Sequence of humanized antibody <220>

<221> CECHA_RÓŻN.

<223> Ab-15 heavy chain <400> 72

VP / 4572 / AGR

EP 2 569 010 B1

<img file="PL2569010T3_D0002.tif" />

VP / 4572 / AGR

EP 2 569 010 B1

40 45

Gly Arg He Asp Pro Glu Asn Gly Asp Thr Leu Tyr Asp Pro Lys Phe 50 55 60

Gin Asp Lys Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Leu Arg Ser Asp Asp Thr Al Val Tyr Tyr Cys 85 90 95

Ala Arg Glu Ala Asp Tyr Phe His Asp Gly Thr Ser Tyr Trp Tyr Phe 100 105 110

Asp Val Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 115 120 125

Lys Gly Pro Cheese Val Phe Pro Leu Ala Pro Cys Ser Arg Cheese Thr Cheese 130 135 140

Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Ly3 Asp Tyr Phe Pro Glu 145 150 155 160

Pro Val Thr Val Ser Trp A3n Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175

Thr Phe Pro Ala Val Leu Gin Cheese Cheese Gly Leu Tyr Cheese Leu Cheese Ser 180 185 190

Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gin Thr Tyr Thr Cys 195 200 205

Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu 210 215 220

Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Al Pro Pro Val Ala 225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270

Glu Asp Pro Glu Val Gin Phe Asn Trp Tyr Val Asp Gly Val Glu Val

VP / 4572 / AGR

EP 2 569 010 B1

275, 280,285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Phe Asn Ser Thr Phe 290 295 300

Val Val Val Ser Val Leu Thr Val Val His Gin Asp Trp Leu Asn Gly 305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile 325 330 335

Glu Lys Thr Ile Ser Ly3 Thr Lys Gly Gin Pro Arg Glu Pro Gin Val 340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Met Thr Lys Asn Gin Val Ser 355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Cheese Asp Ile Al Val Glu 370 375 380

Trp Glu Cheese Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400

Met Leu Asp Ser Asp Gly Cheese Phe Phe Leu Tyr Cheese Lys Leu Thr Val 405 410 415

Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Cheese 435 440 445

Pro Gly Lys 450 <210> 73 <211> 11 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-4 and Ab-5 <400> 73

Arg Ala Ser Gin Asp Ile Ser Asn Tyr Leu Asn 15 10 <210> 74 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-4 and Ab-5 <400> 74

Tyr Thr Ser Arg Leu Ser 1 5 <210> 75 <211> 9

VP / 4572 / AGR

EP 2 569 010 B1 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-4 and Ab-5 <400> 75

Gin Gin Glyl A3p Thr Leu Pro Tyr Thr 1 5 <210> 76 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-4 and Ab-5 <400> 76

Asp Tyr Asn Met His 1 5 <210> 77 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-4 and Ab-5 <400> 77

Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr A3n Gin Lys Phe Lys 15 10 15

Gly <210> 78 <211> 14 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 <400> 78

Leu Gly Tyr Asp Asp. Aspr. Asp Asp. Trp Tyr Phe Asp. Val. 1 5 10 <210> 79 <211> 645 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-4 light chain <400> 79

VP / 4572 / AGR

EP 2 569 010 B1

<td>gatatccaga</td><td>tgacacagat</td><td>tacatcctcc</td><td>ctgtctgcct</td><td>ctctgggaga</td><td>cagggtctcc</td><td>60</td>

<td>atcagttgca</td><td>gggcaagtca</td><td>agacattagc</td><td>aattatttaa</td><td>actggtatca</td><td>gcagaaacca</td><td>120</td>

<td>gatggaactt</td><td>ttaaactcct</td><td>tatcttctac</td><td>acatcaagat</td><td>tactctcagg</td><td>agtcccatca</td><td>180</td>

<td>aggttcagtg</td><td>gcagtgggtc</td><td>tggaacagat</td><td>tattctctca</td><td>ccatttacaa</td><td>cctggagcaa</td><td>240</td>

<td>gaagattttg</td><td>ccacttactt</td><td>ttgccaacag</td><td>ggagatacgc</td><td>ttccgtacac</td><td>tttcggaggg</td><td>300</td>

<td>gggaccaagc</td><td>tggaaataaa</td><td>acgggctgat</td><td>gctgcaccaa</td><td>ctgtatccat</td><td>cttcccacca</td><td>360</td>

<td>tccagtgagc</td><td>agttaacatc</td><td>tggaggtgcc</td><td>tcagtcgtgt</td><td>gcttcttgaa</td><td>caacttctac</td><td>420</td>

<td>cccaaagaca</td><td>tcaatgtcaa</td><td>gtggaagatt</td><td>gatggcagtg</td><td>aacgacaaaa</td><td>tggcgtcctg</td><td>480</td>

<td>aacagttgga</td><td>ctgatcagga</td><td>cagcaaagac</td><td>agcacctaca</td><td>gcatgagcag</td><td>caccctcacg</td><td>540</td>

<td>ttgaccaagg</td><td>acgagtatga</td><td>acgacataac</td><td>agctatacct</td><td>gtgaggccac</td><td>tcacaagaca</td><td>600</td>

<td>tcaacttcac</td><td>ccattgtcaa</td><td>gagcttcaac</td><td>aggaatgagt</td><td>gttag</td><td></td><td>645</td>

<210> 80 <211> 214 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN. <223> Ab-4 light chain <400> 80

VP / 4572 / AGR

EP 2 569 010 B1

Asp Ile Gin Met Thr Gin Ile Thr Ser Ser Leu Ser Ala Ser Leu Gly 15 10 15

Asp Arg. Val Ser. Ser. Ser Cys. Arg. Ala. Ser Gin. Asp. Ser Asn. Tyr 20 25 30

Leu Asn Trp Tyr Gin Gin Lys Pro Asp Gly Thr Phe Lys Leu Leu Ile 35 40 45

Phe Tyr Thr Ser Arg Leu Leu Cheese Gly Val Pro Cheese Arg Phe Cheese Gly 50 55 60

Cheese Gly Cheese Gly Thr Asp Tyr Cheese Leu Thr Ile Tyr Asn Leu Glu Gin 65 70 75 80

Glu Asp Phe Ala Thr Tyr Phe Cys Gin Gin Gly Asp Thr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110

Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gin Leu Thr Cheese Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140

Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu 145 150 155 160

Asn Ser Trp Thr Asp Gin Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Cheese Tyr 180 185 190

Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205

Phe Asn Arg Asn Glu Cy3 210 <210> 81 <211> 1344 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-4 heavy chain <400> 81

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggtccaac</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctaatgaagc</td><td>ctggggcttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggata</td><td>tacattcact</td><td>gactacaaca</td><td>tgcactgggt</td><td>gaagcagaac</td><td>120</td>

<td>caaggaaaga</td><td>ccctagagtg</td><td>gataggagaa</td><td>attaatccta</td><td>acagtggtgg</td><td>tgctggctac</td><td>180</td>

<td>aaccagaagt</td><td>tcaagggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcctccac</td><td>cacagcctac</td><td>240</td>

<td>atggagctcc</td><td>gcagcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagattgggc</td><td>300</td>

<td>tacgatgata</td><td>tctacgacga</td><td>ctggtacttc</td><td>gatgtctggg</td><td>gcgcagggac</td><td>cacggtcacc</td><td>360</td>

<td>gtctcctcag</td><td>ccaaaacgac</td><td>acccccatct</td><td>gtctatccac</td><td>tggcccctgg</td><td>atctgctgcc</td><td>420</td>

<td>caaactaact</td><td>ccatggtgac</td><td>cctgggatgc</td><td>ctggtcaagg</td><td>gctatttccc</td><td>tgagccagtg</td><td>480</td>

<td>acagtgacct</td><td>ggaactctgg</td><td>atccctgtcc</td><td>agcggtgtgc</td><td>acaccttccc</td><td>agctgtcctg</td><td>540</td>

<td>cagtctgacc</td><td>tctacactct</td><td>gagcagctca</td><td>gtgactgtcc</td><td>cctccagcac</td><td>ctggcccagc</td><td>600</td>

<td>gagaccgtca</td><td>cctgcaacgt</td><td>tgcccacccg</td><td>gccagcagca</td><td>ccaaggtgga</td><td>caagaaaatt</td><td>660</td>

<td>gtgcccaggg</td><td>attgtggttg</td><td>taagccttgc</td><td>atatgtacag</td><td>tcccagaagt</td><td>atcatctgtc</td><td>720</td>

<td>ttcatcttcc</td><td>ccccaaagcc</td><td>caaggatgtg</td><td>ctcaccatta</td><td>ctctgactcc</td><td>taaggtcacg</td><td>780</td>

<td>tgtgttgtgg</td><td>tagacatcag</td><td>caaggatgat</td><td>cccgaggtcc</td><td>agttcagctg</td><td>gtttgtagat</td><td>840</td>

<td>gatgtggagg</td><td>tgcacacagc</td><td>tcagacgcaa</td><td>ccccgggagg</td><td>agcagttcaa</td><td>cagcactttc</td><td>900</td>

<td>cgctcagtca</td><td>gtgaacttcc</td><td>catcatgcac</td><td>caggactggc</td><td>tcaatggcaa</td><td>ggagttcaaa</td><td>960</td>

<td>tgcagggtca</td><td>acagtgcagc</td><td>tttccctgcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>ggcagaccga</td><td>aggctccaca</td><td>ggtgtacacc</td><td>attccacctc</td><td>ccaaggagca</td><td>gatggccaag</td><td>1080</td>

<td>gataaagtca</td><td>gtctgacctg</td><td>catgataaca</td><td>gacttcttcc</td><td>ctgaagacat</td><td>tactgtggag</td><td>1140</td>

<td>tggcagtgga</td><td>atgggcagcc</td><td>agcggagaac</td><td>tacaagaaca</td><td>ctcagcccat</td><td>catggacaca</td><td>1200</td>

<td>gatggctctt</td><td>acttcatcta</td><td>cagcaagctc</td><td>aatgtgcaga</td><td>agagcaactg</td><td>ggaggcagga</td><td>1260</td>

<td>aatactttca</td><td>cctgctctgt</td><td>gttacatgag</td><td>ggcctgcaca</td><td>accaccatac</td><td>tgagaagagc</td><td>1320</td>

<td>ctctcccact</td><td>ctcctggtaa</td><td>atga</td><td></td><td></td><td></td><td>1344</td>

<210> 82 <211> 447 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-4 heavy chain <400> 82

VP / 4572 / AGR

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cy3 Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Asn Met His Trp Val Lys Gin Asn Gin Gly Lys Thr Leu Glu Trp Ile 35 40 45

Gly Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr Asn Gin Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Thr Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Gly Tyr Asp Asp. Ile Asp. Asp Asp Trp Tyr Phe Asp Val 100 105 110

Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Al Lys Thr Thr Pro 115 120 125

Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr Asn Cheese 130 135 140

Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val 145 150 155 160

Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe 165 170 175

Pro Ala Val Leu Gin Cheese Asp Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr 180 185 190

Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala 195 200 205

His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp 210 215 220

Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val

VP / 4572 / AGR

EP 2 569 010 B1

225 230 235 240

Phe Ile Phe Pro Pro Lye Pro Lys Asp Val Leu Thr Ile Thr Leu Thr 245 250 255

Pro Lys Val Thr Cys Val Val Val Asp. Ile Ser Lys Asp Asp Pro Glu 260 265 270

Val Gin Phe Ser Trp Phe Val Asp Asp. Val Gl. Val. His Thr. Ala Gin 275 280 285

Thr Gin Pro Arg Glu Glu Gin Phe Asn Cheese Thr Phe Arg Cheese Val Ser 290 295 300

Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Lys 305 310 315 320

Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Ile Glu Lys Thr Ile 325 330 335

Ser Lye Thr Lys Gly Arg Pro Lys Al Pro Gin Val Tyr Thr Ile Pro 340 345 350

Pro Pro Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met 355 360 365

How much Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn 370 375 380

Gly Gin Pro Ala Glu Asn Tyr Lys A3n Thr Gin Pro How Much Met Asp Thr 385 390 395 400

A3p Gly Cheese Tyr Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys Cheese Asn 405 410 415

Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu 420 425 430

His Asn His Thr Glu Lys Ser Leu Ser His Cheese Pro Gly Lys 435 440 445 <210> 83 <211> 321 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Lightweight variable Ab-5 <400> 83

VP / 4572 / AGR

EP 2 569 010 B1

<td>gacatccaga</td><td>tgacccagtc</td><td>tccatcctcc</td><td>ctctccgcat</td><td>ccgtaggcga</td><td>ccgcgtaacc</td><td>60</td>

<td>ataacatgta</td><td>gagcatctca</td><td>agatatttcc</td><td>aactatttga</td><td>attggtacca</td><td>acaaaaaccc</td><td>120</td>

<td>ggcaaagcac</td><td>ctaaactcct</td><td>catttactat</td><td>acatcaagac</td><td>tcctctccgg</td><td>cgttccatca</td><td>180</td>

<td>cgattctcag</td><td>gctccggctc</td><td>cggcacagat</td><td>ttcacactca</td><td>ctatttcctc</td><td>cctccaacca</td><td>240</td>

<td>gaagattttg</td><td>caacctatta</td><td>ctgtcaacaa</td><td>ggcgatacac</td><td>tcccatacac</td><td>attcggcggc</td><td>300</td>

<td>ggcacaaaag</td><td>ttgaaattaa</td><td>and</td><td></td><td></td><td></td><td>321</td>

<210> 84 <211> 107 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Lightweight variable Ab-5

<td colspan="5">&lt;400&gt; 84</td><td colspan="2" rowspan="2">Gin Cheese</td><td colspan="2" rowspan="2">Pro Ser</td><td rowspan="2">Cheese 10</td><td rowspan="2">Leu</td><td rowspan="2">Cheese</td><td rowspan="2">ala</td><td rowspan="2">Cheese</td><td rowspan="2">val 15</td><td rowspan="2">Gly</td>

<td>Asp 1</td><td colspan="3">Ile Gin Met</td><td>Thr 5</td>

<td>Asp</td><td>Arg</td><td>val</td><td>Thr</td><td>How much</td><td>Thr</td><td>Cy3</td><td>Arg</td><td>ala</td><td>Cheese</td><td>Gin</td><td>Asp</td><td>How much</td><td>Cheese</td><td>own</td><td>Tyr</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>Leu</td><td>own</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>lys</td><td>Pro</td><td>Gly</td><td>Lye</td><td>ala</td><td>Pro</td><td>lys</td><td>Leu</td><td>Leu</td><td>How much</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Tyr</td><td>Tyr</td><td>Thr</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>Leu</td><td>Cheese</td><td>Gly</td><td>val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>phe</td><td>Cheese</td><td>Gly</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td><td>Pro</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Glu</td><td>Asp</td><td>phe</td><td>ala</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Pro</td><td>Tyr</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>Thr</td><td>phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>lys</td><td>val</td><td>Glu</td><td>How much</td><td>Ly3</td><td></td><td></td><td></td><td></td><td></td>

100 105 <210> 85 <211> 369 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Heavy variable Ab-5 <400> 85

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggtgcagc</td><td>tggtgcagag</td><td>cggcgccgag</td><td>gtaaaaaaac</td><td>caggagcaag</td><td>cgttaaagtt</td><td>60</td>

<td>tcttgtaaag</td><td>caagcggata</td><td>tacatttaca</td><td>gattacaaca</td><td>tgcattgggt</td><td>aagacaagcg</td><td>120</td>

<td>ccaggacaag</td><td>gattggaatg</td><td>gatgggcgaa</td><td>attaacccta</td><td>atagtggagg</td><td>agcaggctac</td><td>180</td>

<td>aatcaaaaat</td><td>tcaaagggag</td><td>agttacaatg</td><td>acaacagaca</td><td>caagcacttc</td><td>aacagcatat</td><td>240</td>

<td>atggaactgc</td><td>gatcacttag</td><td>aagcgacgat</td><td>acagctgtat</td><td>actattgcgc</td><td>acgacttggg</td><td>300</td>

<td>tatgatgata</td><td>tatatgatga</td><td>ctggtatttc</td><td>gatgtttggg</td><td>gccagggaac</td><td>aacagttacc</td><td>360</td>

gtctctagt 369 <210> 86 <211> 123 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Heavy variable Ab-5 <400> 86

Glu Val Gin Leu Val Gin Cheese Gly Ala Glu Val Lys Pro Gly Ala 15 10 15

Ser Val Lys Val Ser Cy3 Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Asn Met His Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr Asn Gin Lys Phe 50 55 60

Lys Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Leu Arg Ser Asp Asp Thr Al Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Gly Tyr Asp Asp. Ile Asp. Asp Asp Trp Tyr Phe Asp Val 100 105 110

Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 87 <211> 642 <212> DNA <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> trait_different

<223> Ab-5 light chain <400> 87

VP / 4572 / AGR

EP 2 569 010 B1

<td>gacatccaga</td><td>tgacccagtc</td><td>tccatcctcc</td><td>ctctccgcat</td><td>ccgtaggcga</td><td>ccgcgtaacc</td><td>60</td>

<td>ataacatgta</td><td>gagcatctca</td><td>agatatttcc</td><td>aactatttga</td><td>attggtacca</td><td>acaaaaaccc</td><td>120</td>

<td>ggcaaagcac</td><td>ctaaactcct</td><td>catttactat</td><td>acatcaagac</td><td>tcctctccgg</td><td>cgttccatca</td><td>180</td>

<td>cgattctcag</td><td>gctccggctc</td><td>cggcacagat</td><td>ttcacactca</td><td>ctatttcctc</td><td>cctccaacca</td><td>240</td>

<td>gaagattttg</td><td>caacctatta</td><td>ctgtcaacaa</td><td>ggcgatacac</td><td>tcccatacac</td><td>attcggcggc</td><td>300</td>

<td>ggcacaaaag</td><td>ttgaaattaa</td><td>acgtacggtg</td><td>gctgcaccat</td><td>ctgtcttcat</td><td>cttcccgcca</td><td>360</td>

<td>tctgatgagc</td><td>agttgaaatc</td><td>tggaactgcc</td><td>tctgttgtgt</td><td>gcctgctgaa</td><td>taacttctat</td><td>420</td>

<td>cccagagagg</td><td>ccaaagtaca</td><td>gtggaaggtg</td><td>gataacgccc</td><td>tccaatcggg</td><td>taactcccag</td><td>480</td>

<td>gagagtgtca</td><td>cagagcagga</td><td>cagcaaggac</td><td>agcacctaca</td><td>gcctcagcag</td><td>caccctgacg</td><td>540</td>

<td>ctgagcaaag</td><td>cagactacga</td><td>gaaacacaaa</td><td>gtctacgcct</td><td>gcgaagtcac</td><td>ccatcagggc</td><td>600</td>

<td>ctgagctcgc</td><td>ccgtcacaaa</td><td>gagcttcaac</td><td>aggggagagt</td><td>gt</td><td></td><td>642</td>

<210> 88 <211> 214 <212> PRT <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> CECHA_RÓŻN.

<223> Ab-5 light chain <400> 88

VP / 4572 / AGR

EP 2 569 010 B1

Asp Ile Gin Met Thr Gin Cheese Pro Ser Ser Leu Cheese Ala Cheese Val Gly 15 10 15

A3p Arg Val Thr Ile Thr Cys Arg Ala Ser Gin Asp Ile Ser Asn Tyr 20 25 30

Leu Asn Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45

Tyr Tyr Thr Ser Arg Leu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Cheese Gly Cheese Gly Thr Asp Phe Thr Leu Thr Ile Cheese Cheese Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Gly Asp Thr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe Ile Phe Pro Pro. Ser. Glu Gin Leu Lys Cheese Gly 115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Cheese Gly Asn Cheese Gin 145 150 155 160

Glu Cheese Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Cheese 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Cheese Ser Pro Val Thr Lys Ser 195 200 205

Phe Asn Arg Gly Glu Cys 210 <210> 89 <211> 1347 <212> DNA <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> trait_different

<223> Ab-5 heavy chain <400> 89

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggtgcagc</td><td>tggtgcagag</td><td>cggcgccgag</td><td>gtaaaaaaac</td><td>caggagcaag</td><td>cgttaaagtt</td><td>60</td>

<td>tcttgtaaag</td><td>caagcggata</td><td>tacatttaca</td><td>gattacaaca</td><td>tgcattgggt</td><td>aagacaagcg</td><td>120</td>

<td>ccaggacaag</td><td>gattggaatg</td><td>gatgggcgaa</td><td>attaacccta</td><td>atagtggagg</td><td>agcaggctac</td><td>180</td>

<td>aatcaaaaat</td><td>tcaaagggag</td><td>agttacaatg</td><td>acaacagaca</td><td>caagcacttc</td><td>aacagcatat</td><td>240</td>

<td>atggaactgc</td><td>gatcacttag</td><td>aagcgacgat</td><td>acagctgtat</td><td>actattgcgc</td><td>acgacttggg</td><td>300</td>

<td>tatgatgata</td><td>tatatgatga</td><td>ctggtatttc</td><td>gatgtttggg</td><td>gccagggaac</td><td>aacagttacc</td><td>360</td>

<td>gtctctagtg</td><td>cctccaccaa</td><td>gggcccatcg</td><td>gtcttccccc</td><td>tggcgccctg</td><td>ctccaggagc</td><td>420</td>

<td>acctccgaga</td><td>gcacagcggc</td><td>cctgggctgc</td><td>ctggtcaagg</td><td>actacttccc</td><td>cgaaccggtg</td><td>480</td>

<td>acggtgtcgt</td><td>ggaactcagg</td><td>cgctctgacc</td><td>agcggcgtgc</td><td>acaccttccc</td><td>agctgtccta</td><td>540</td>

<td>cagtcctcag</td><td>gactctactc</td><td>cctcagcagc</td><td>gtggtgaccg</td><td>tgccctccag</td><td>caacttcggc</td><td>600</td>

<td>acccagacct</td><td>acacctgcaa</td><td>cgtagatcac</td><td>aagcccagca</td><td>acaccaaggt</td><td>ggacaagaca</td><td>660</td>

<td>gttgagcgca</td><td>aatgttgtgt</td><td>cgagtgccca</td><td>ccgtgcccag</td><td>caccacctgt</td><td>ggcaggaccg</td><td>720</td>

<td>tcagtcttcc</td><td>tcttcccccc</td><td>aaaacccaag</td><td>gacaccctca</td><td>tgatctcccg</td><td>gacccctgag</td><td>780</td>

<td>gtcacgtgcg</td><td>tggtggtgga</td><td>cgtgagccac</td><td>gaagaccccg</td><td>aggtccagtt</td><td>caactggtac</td><td>840</td>

<td>gtggacggcg</td><td>tggaggtgca</td><td>taatgccaag</td><td>acaaagccac</td><td>gggaggagca</td><td>gttcaacagc</td><td>900</td>

<td>acgttccgtg</td><td>tggtcagcgt</td><td>cctcaccgtt</td><td>gtgcaccagg</td><td>actggctgaa</td><td>cggcaaggag</td><td>960</td>

<td>tacaagtgca</td><td>aggtctccaa</td><td>caaaggcctc</td><td>ccagccccca</td><td>tcgagaaaac</td><td>catctccaaa</td><td>1020</td>

<td>accaaagggc</td><td>agccccgaga</td><td>accacaggtg</td><td>tacaccctgc</td><td>ccccatcccg</td><td>ggaggagatg</td><td>1080</td>

<td>accaagaacc</td><td>aggtcagcct</td><td>gacctgcctg</td><td>gtcaaaggct</td><td>tctaccccag</td><td>cgacatcgcc</td><td>1140</td>

<td>gtggagtggg</td><td>agagcaatgg</td><td>gcagccggag</td><td>aacaactaca</td><td>agaccacacc</td><td>tcccatgctg</td><td>1200</td>

<td>gactccgacg</td><td>gctccttctt</td><td>cctctacagc</td><td>aagctcaccg</td><td>tggacaagag</td><td>caggtggcag</td><td>1260</td>

<td>caggggaacg</td><td>tcttctcatg</td><td>ctccgtgatg</td><td>catgaggctc</td><td>tgcacaacca</td><td>ctacacgcag</td><td>1320</td>

<td>aagagcctct</td><td>ccctgtctcc</td><td>gggtaaa</td><td></td><td></td><td></td><td>1347</td>

<210> 90 <211> 449 <212> PRT <213> Artificial sequence <220>

<223> Humanized antibody <220>

<221> CECHA_RÓŻN.

<223> Ab-5 heavy chain <400> 90

VP / 4572 / AGR

EP 2 569 010 B1

Glu Val Gin Leu Val Gin Cheese Gly Ala Glu Val Lys Pro Gly Ala 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Asn Met His Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr Asn Gin Ly3 Phe 50 55 60

Lye Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Leu Arg Ser Asp Asp Thr Al Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Gly Tyr Asp Asp. Ile Asp. Asp Asp Trp Tyr Phe Asp Val 100 105 110

Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ser Ser Thr Lys Gly 115 120 125

Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Cheese 130 135 140

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175

Pro Ala Val Leu Gin Cheese Cheese Gly Leu Tyr Cheese Leu Cheese Cheese Val Val 180 185 190

Thr Val Pro Ser Ser Asn Phe Gly Thr Gin Thr Tyr Thr Cys Asn Val 195 200 205

Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys

VP / 4572 / AGR

EP 2 569 010 B1

<td></td><td>210</td><td></td><td></td><td></td><td></td><td>215</td><td></td><td></td><td></td><td></td><td>220</td><td></td><td></td><td></td><td></td>

<td>Cye</td><td>Cy3</td><td>val</td><td>Glu</td><td>Cy3</td><td>Pro</td><td>Pro</td><td>Cys</td><td>Pro</td><td>ala</td><td>Pro</td><td>Pro</td><td>val</td><td>ala</td><td>Gly</td><td>Pro</td>

<td>225</td><td></td><td></td><td></td><td></td><td>230</td><td></td><td></td><td></td><td></td><td>235</td><td></td><td></td><td></td><td></td><td>240</td>

<td>Cheese</td><td>val</td><td>phe</td><td>Leu</td><td>phe</td><td>Pro</td><td>Pro</td><td>Ly3</td><td>Pro</td><td>lys</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Underworld</td><td>How much</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td>245</td><td></td><td></td><td></td><td></td><td>250</td><td></td><td></td><td></td><td></td><td>255</td><td></td>

<td>Arg</td><td>Thr</td><td>Pro</td><td>Glu</td><td>val</td><td>Thr</td><td>Cye</td><td>val</td><td>val</td><td>val</td><td>Asp</td><td>val</td><td>Cheese</td><td>His</td><td>Glu</td><td>Asp</td>

<td></td><td></td><td></td><td>260</td><td></td><td></td><td></td><td></td><td>265</td><td></td><td></td><td></td><td></td><td>270</td><td></td><td></td>

<td>Pro</td><td>Glu</td><td>val</td><td>Gin</td><td>phe</td><td>own</td><td>Trp</td><td>Tyr</td><td>val</td><td>Asp</td><td>Gly</td><td>val</td><td>Glu</td><td>val</td><td>His</td><td>own</td>

<td></td><td></td><td>275</td><td></td><td></td><td></td><td></td><td>280</td><td></td><td></td><td></td><td></td><td>285</td><td></td><td></td><td></td>

<td>ala</td><td>lys</td><td>Thr</td><td>lys</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>phe</td><td>own</td><td>Cheese</td><td>Thr</td><td>phe</td><td>Arg</td><td>val</td>

<td></td><td>290</td><td></td><td></td><td></td><td></td><td>295</td><td></td><td></td><td></td><td></td><td>300</td><td></td><td></td><td></td><td></td>

<td>val</td><td>Cheese</td><td>val</td><td>Leu</td><td>Thr</td><td>val</td><td>val</td><td>His</td><td>Gin</td><td>Asp</td><td>Trp</td><td>Leu</td><td>own</td><td>Gly</td><td>lys</td><td>Glu</td>

<td>305</td><td></td><td></td><td></td><td></td><td>310</td><td></td><td></td><td></td><td></td><td>315</td><td></td><td></td><td></td><td></td><td>320</td>

<td>Tyr</td><td>Ly3</td><td>Cys</td><td>lys</td><td>val</td><td>Cheese</td><td>own</td><td>lys</td><td>Gly</td><td>Leu</td><td>Pro</td><td>ala</td><td>Pro</td><td>How much</td><td>Glu</td><td>lys</td>

<td></td><td></td><td></td><td></td><td>325</td><td></td><td></td><td></td><td></td><td>330</td><td></td><td></td><td></td><td></td><td>335</td><td></td>

<td>Thr</td><td>How much</td><td>Cheese</td><td>lys</td><td>Thr</td><td>lys</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Pro</td><td>Gin</td><td>val</td><td>Tyr</td><td>Thr</td>

<td></td><td></td><td></td><td>340</td><td></td><td></td><td></td><td></td><td>345</td><td></td><td></td><td></td><td></td><td>350</td><td></td><td></td>

<td>Leu</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Underworld</td><td>Thr</td><td>lys</td><td>A3n</td><td>Gin</td><td>val</td><td>Cheese</td><td>Leu</td><td>Thr</td>

<td></td><td></td><td>355</td><td></td><td></td><td></td><td></td><td>360</td><td></td><td></td><td></td><td></td><td>365</td><td></td><td></td><td></td>

<td>Cys</td><td>Leu</td><td>val</td><td>lys</td><td>Gly</td><td>phe</td><td>Tyr</td><td>Pro</td><td>Cheese</td><td>Asp</td><td>How much</td><td>ala</td><td>val</td><td>Glu</td><td>Trp</td><td>Glu</td>

<td></td><td>370</td><td></td><td></td><td></td><td></td><td>375</td><td></td><td></td><td></td><td></td><td>380</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>own</td><td>Gly</td><td>Gin</td><td>Pro</td><td>Glu</td><td>own</td><td>own</td><td>Tyr</td><td>lys</td><td>Thr</td><td>Thr</td><td>Pro</td><td>Pro</td><td>Underworld</td><td>Leu</td>

<td>385</td><td></td><td></td><td></td><td></td><td>390</td><td></td><td></td><td></td><td></td><td>395</td><td></td><td></td><td></td><td></td><td>400</td>

<td>Asp</td><td>Cheese</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>phe</td><td>phe</td><td>Leu</td><td>Tyr</td><td>Cheese</td><td>Ly3</td><td>Leu</td><td>Thr</td><td>val</td><td>Asp</td><td>Lye</td>

<td></td><td></td><td></td><td></td><td>405</td><td></td><td></td><td></td><td></td><td>410</td><td></td><td></td><td></td><td></td><td>415</td><td></td>

<td>Cheese</td><td>Arg</td><td>Trp</td><td>Gin</td><td>Gin</td><td>Gly</td><td>own</td><td>val</td><td>phe</td><td>Cheese</td><td>Cy3</td><td>Cheese</td><td>val</td><td>Underworld</td><td>His</td><td>Glu</td>

<td></td><td></td><td></td><td>420</td><td></td><td></td><td></td><td></td><td>425</td><td></td><td></td><td></td><td></td><td>430</td><td></td><td></td>

<td>ala</td><td>Leu</td><td>His</td><td>own</td><td>His</td><td>Tyr</td><td>Thr</td><td>Gin</td><td>lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Pro</td><td>Gly</td>

<td></td><td></td><td>435</td><td></td><td></td><td></td><td></td><td>440</td><td></td><td></td><td></td><td></td><td>445</td><td></td><td></td><td></td>

Lys <210> 91 <211> 11 <212> PRT <213> Artificial sequence <220>

VP / 4572 / AGR

EP 2 569 010 B1 <223> LCDR1 Ab-6 <400> 91

Arg Ala Ser Gin Asp 1 5 <210> 92 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-6 <400> 92

Tyr Thr Ser Arg Leu His 1 5 <210> 93 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-6 <400> 93

Gin Gin Gly Asp Thr Leu Pro Tyr Thr

5 <210> 94 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-6 <400> 94

Asp Tyr A3n Met His

5 <210> 95 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-6 <400> 95

Ile Ser Αβη Tyr

Cheese

Leu

own

<img file="PL2569010T3_D0003.tif" />

<210> 96 <211> 14 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-6 <400> 96

Leu Val Tyr Asp Gly Cheese Tyr Glu Asp

5 <210> 97 <211> 645 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-6 light chain <400> 97

Trp

Tyr Phe Asp Val

VP / 4572 / AGR

EP 2 569 010 B1

<td>gatatccaga</td><td>tgacacagac</td><td>tacatcctcc</td><td>ctgtctgcct</td><td>ctctgggaga</td><td>cagagtcacc</td><td>60</td>

<td>atcagttgca</td><td>gggcaagtca</td><td>ggacattagc</td><td>aattatttaa</td><td>actggtttca</td><td>gcagaaacca</td><td>120</td>

<td>gatggaactc</td><td>ttaaactcct</td><td>gatcttctac</td><td>acatcaagat</td><td>tacactcagg</td><td>agttccatca</td><td>180</td>

<td>aggttcagtg</td><td>gcagtgggtc</td><td>tggaacagat</td><td>tattctctca</td><td>ccattagcaa</td><td>cctggagcaa</td><td>240</td>

<td>gaagatattg</td><td>ccacttactt</td><td>ttgccaacag</td><td>ggtgatacgc</td><td>ttccgtacac</td><td>gttcgggggg</td><td>300</td>

<td>gggaccaagc</td><td>tggaaataag</td><td>acgggctgat</td><td>gctgcaccaa</td><td>ctgtatccat</td><td>cttcccacca</td><td>360</td>

<td>tccagtgagc</td><td>agttaacatc</td><td>tggaggtgcc</td><td>tcagtcgtgt</td><td>gcttcttgaa</td><td>caacttctac</td><td>420</td>

<td>cccaaagaca</td><td>tcaatgtcaa</td><td>gtggaagatt</td><td>gatggcagtg</td><td>aacgacaaaa</td><td>tggcgtcctg</td><td>480</td>

<td>aacagttgga</td><td>ctgatcagga</td><td>cagcaaagac</td><td>agcacctaca</td><td>gcatgagcag</td><td>caccctcacg</td><td>540</td>

<td>ttgaccaagg</td><td>acgagtatga</td><td>acgacataac</td><td>agctatacct</td><td>gtgaggccac</td><td>tcacaagaca</td><td>600</td>

<td>tcaacttcac</td><td>ccattgtcaa</td><td>gagcttcaac</td><td>aggaatgagt</td><td>gttag</td><td></td><td>645</td>

<210> 98 <211> 214 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN. <223> Ab-6 light chain <400> 98

VP / 4572 / AGR

EP 2 569 010 B1

Asp Ile Gin Met Thr Gin Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 15 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gin Asp Ile Ser Asn Tyr 20 25 30

Leu Asn Trp Phe Gin Gin Lys Pro Asp Gly Thr Leu Lys Leu Leu Ile 35 40 45

Phe Tyr Thr Ser Arg Leu His Cheese Gly Val Pro Ser Arg Phe Cheese Gly 50 55 60

Cheese Gly Cheese Gly Thr Asp Tyr Cheese Leu Thr Ile Cheese Asn Leu Glu Gin 65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gin Gin Gly Asp Thr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Arg Arg Ala Asp Ala Ala 100 105 110

Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gin Leu Thr Cheese Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140

Asn Val Lys Trp Ly3 Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu 145 150 155 160

Asn Ser Trp Thr Asp Gin Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His A3n Cheese Tyr 180 185 190

Thr Cys Glu Ala Thr Hi3 Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205

Phe Asn Arg Asn Glu Cy3 210 <210> 99 <211> 1344 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-6 heavy chain <400> 99

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggtccagc</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctaatgaagc</td><td>ctggggcttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggata</td><td>cacattcact</td><td>gactacaaca</td><td>tgcactgggt</td><td>gaaacagaac</td><td>120</td>

<td>caaggaaaga</td><td>gcctagagtg</td><td>gataggagaa</td><td>attaatccta</td><td>acagtggtgg</td><td>tagtggctac</td><td>180</td>

<td>aaccaaaagt</td><td>tcaaaggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcttccag</td><td>cacagcctac</td><td>240</td>

<td>atggagctcc</td><td>gcagcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagattggtc</td><td>300</td>

<td>tacgatggca</td><td>gctacgagga</td><td>ctggtacttc</td><td>gatgtctggg</td><td>gcgcagggac</td><td>cacggtcacc</td><td>360</td>

<td>gtctcctcag</td><td>ccaaaacgac</td><td>acccccatct</td><td>gtctatccac</td><td>tggcccctgg</td><td>atctgctgcc</td><td>420</td>

<td>caaactaact</td><td>ccatggtgac</td><td>cctgggatgc</td><td>ctggtcaagg</td><td>gctatttccc</td><td>tgagccagtg</td><td>480</td>

<td>acagtgacct</td><td>ggaactctgg</td><td>atccctgtcc</td><td>agcggtgtgc</td><td>acaccttccc</td><td>agctgtcctg</td><td>540</td>

<td>cagtctgacc</td><td>tctacactct</td><td>gagcagctca</td><td>gtgactgtcc</td><td>cctccagcac</td><td>ctggcccagc</td><td>600</td>

<td>gagaccgtca</td><td>cctgcaacgt</td><td>tgcccacccg</td><td>gccagcagca</td><td>ccaaggtgga</td><td>caagaaaatt</td><td>660</td>

<td>gtgcccaggg</td><td>attgtggttg</td><td>taagccttgc</td><td>atatgtacag</td><td>tcccagaagt</td><td>atcatctgtc</td><td>720</td>

<td>ttcatcttcc</td><td>ccccaaagcc</td><td>caaggatgtg</td><td>ctcaccatta</td><td>ctctgactcc</td><td>taaggtcacg</td><td>780</td>

<td>tgtgttgtgg</td><td>tagacatcag</td><td>caaggatgat</td><td>cccgaggtcc</td><td>agttcagctg</td><td>gtttgtagat</td><td>840</td>

<td>gatgtggagg</td><td>tgcacacagc</td><td>tcagacgcaa</td><td>ccccgggagg</td><td>agcagttcaa</td><td>cagcactttc</td><td>900</td>

<td>cgctcagtca</td><td>gtgaacttcc</td><td>catcatgcac</td><td>caggactggc</td><td>tcaatggcaa</td><td>ggagttcaaa</td><td>960</td>

<td>tgcagggtca</td><td>acagtgcagc</td><td>tttccctgcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>ggcagaccga</td><td>aggctccaca</td><td>ggtgtacacc</td><td>attccacctc</td><td>ccaaggagca</td><td>gatggccaag</td><td>1080</td>

<td>gataaagtca</td><td>gtctgacctg</td><td>catgataaca</td><td>gacttcttcc</td><td>ctgaagacat</td><td>tactgtggag</td><td>1140</td>

<td>tggcagtgga</td><td>atgggcagcc</td><td>agcggagaac</td><td>tacaagaaca</td><td>ctcagcccat</td><td>catggacaca</td><td>1200</td>

<td>gatggctctt</td><td>acttcatcta</td><td>cagcaagctc</td><td>aatgtgcaga</td><td>agagcaactg</td><td>ggaggcagga</td><td>1260</td>

<td>aatactttca</td><td>cctgctctgt</td><td>gttacatgag</td><td>ggcctgcaca</td><td>accaccatac</td><td>tgagaagagc</td><td>1320</td>

<td>ctctcccact</td><td>ctcctggtaa</td><td>atga</td><td></td><td></td><td></td><td>1344</td>

<210> 100 <211> 447 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN. <223> Ab-6 heavy chain <400> 100

VP / 4572 / AGR

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Asn Met His Trp Val Lys Gin Asn Gin Gly Lys Ser Leu Glu Trp Ile 35 40 45

Gly Glu Ile A3n Pro Asn Cheese Gly Gly Cheese Gly Tyr Asn Gin Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Val Tyr Asp Gly Ser Tyr Glu Asp Trp Tyr Phe Asp Val 100 105 110

Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Al Lys Thr Thr Pro 115 120 125

Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr Asn Cheese 130 135 140

Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val 145 150 155 160

Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe

VP / 4572 / AGR

EP 2 569 010 B1

165 170 175

Pro Ala Val Leu Gin Cheese Asp Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr 180 185 190

Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala 195 200 205

His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp 210 215 220

Cys Gly Cys Ly3 Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val 225 230 235 240

Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr 245 250 255

Pro Lys Val Thr Cys Val Val Val Asp. Ile Ser Lys Asp Asp Pro Glu 260 265 270

Val Gin Phe Ser Trp Phe Val Asp Asp. Val Gl. Val. His Thr. Ala Gin 275 280 285

Thr Gin Pro Arg Glu Glu Gin Phe Asn Cheese Thr Phe Arg Cheese Val Ser 290 295 300

Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Lys 305 310 315 320

Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Ile Glu Lys Thr Ile 325 330 335

Lys Thr Lys Gly Arg Pro Pro Lys Al Pro Gin Val Tyr Thr Ile Pro 340 345 350

Pro Pro Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met 355 360 365

How much Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn 370 375 380

Gly Gin Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro Ile Met Asp Thr 385 390 395 400

Asp Gly Cheese Tyr Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys Cheese Asn

VP / 4572 / AGR

EP 2 569 010 B1

<img file="PL2569010T3_D0004.tif" />

<210> 101 <211> 11 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-7 <400> 101

Arg Ala Ser Gin Val Ile Thr Asn Tyr Leu Tyr 15 10 <210> 102 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-7 <400> 102

Tyr Thr Ser Arg Leu His Ser

5 <210> 103 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-7 <400> 103

Gin Gin Gly Asp Thr Leu Pro Tyr Thr 1 5 <210> 104 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-7 <400> 104

Asp Tyr Asn Met His 1 5 <210> 105 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-7 <400> 105

Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr Asn Gin Gin Phe Lys 15 10 15

Gly <210> 106 <211> 14 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-7

VP / 4572 / AGR

EP 2 569 010 B1 &amp; lt; 400 &amp; gt; 106

Leu Gly Tyr Val Gly Asn Tyr Glu Asp Trp Tyr Phe Asp Val

10 <210> 107 <211> 642 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> light chain of the Ab-7 <400> 107 gatatccaga tgacacagac tacatcctcc ctgtctgcct ctctgggaga cagagtcacc 60 atctgttgca gggcaagtca ggtcattacc aattatttat actggtatca gcagaaacca 120 gatggaactt ttaaactcct gatctactac acatcaagat tacactcagg agtcccatca 160 aggttcagtg gcagtgggtc tggaacagat tattctctca ccattagcaa cctggaacag 240 gaagatattg ccacttactt ttgccaacag ggtgatacgc ttccgtacac gttcggaggg 300 gggaccaagc tggaaataaa acgggctgat gctgcaccaa ctgtatccat cttcccacca 360 tccagtgagc agttaacatc tggaggtgcc tcagtcgtgt gcttcttgaa caacttctac 420 cccaaagaca tcaatgtcaa gtggaagatt gatggcagtg aacgacaaaa tggcgtcctg 480 aacagttgga ctgatcagga cagcaaagac agcacctaca gcatgagcag caccctcacg 540 ttgaccaagg acgagtatga acgacataac agctatacct gtgaggccac tcacaagaca 600 tcaacttcac ccattgtcaa gagcttcaac aggaatgagt gt 642 <210> 108 <211> 214 <212> PRT <213>Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-7 light chain <400> 108

VP / 4572 / AGR

EP 2 569 010 B1

Asp Ile Gin Met Thr Gin Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 15 10 15

Asp Arg Val Thr Ile Cys Cys Arg Ala Ser Gin Val Ile Thr Asn Tyr 20 25 30

Leu Tyr Trp Tyr Gin Gin Lys Pro Asp Gly Thr Phe Lys Leu Leu Ile 35 40 45

Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Cheese Gly Cheese Gly Thr Asp Tyr Cheese Leu Thr Ile Cheese Asn Leu Glu Gin 65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gin Gin G3 Thr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110

Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gin Leu Thr Cheese Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140

Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu 145 150 155 160

Asn Cheese Trp Thr Aep Gin Asp Ser Lys Asp Ser Thr Tyr Cheese Met Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Cheese Tyr 180 185 190

Thr Cy3 Glu Ala Thr Hi3 Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205

Phe A3n Arg Asn Glu Cys 210 <210> 109 <211> 1341 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-7 heavy chain <400> 109

VP / 4572 / AGR

EP 2 569 010 B1

<td>gaggtccagc</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctaatgaagc</td><td>ctggggcttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggata</td><td>cacattcact</td><td>gactacaaca</td><td>tgcactggat</td><td>gaagcagaac</td><td>120</td>

<td>caaggaaaga</td><td>gcctagaatg</td><td>gataggagaa</td><td>attaatccta</td><td>acagtggtgg</td><td>tgctggctac</td><td>180</td>

<td>aaccagcagt</td><td>tcaaaggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcctccag</td><td>gacagcctac</td><td>240</td>

<td>atggagctcc</td><td>gcagcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagattgggc</td><td>300</td>

<td>tacgttggta</td><td>attacgagga</td><td>ctggtacttc</td><td>gatgtctggg</td><td>gcgcagggac</td><td>cacggtcacc</td><td>360</td>

<td>gtctcctcag</td><td>ccaaaacgac</td><td>acccccatct</td><td>gtctatccac</td><td>tggcccctgg</td><td>atctgctgcc</td><td>420</td>

<td>caaactaact</td><td>ccatggtgac</td><td>cctgggatgc</td><td>ctggtcaagg</td><td>gctatttccc</td><td>tgagccagtg</td><td>480</td>

<td>acagtgacct</td><td>ggaactctgg</td><td>atccctgtcc</td><td>agcggtgtgc</td><td>acaccttccc</td><td>agctgtcctg</td><td>540</td>

<td>cagtctgacc</td><td>tctacactct</td><td>gagcagctca</td><td>gtgactgtcc</td><td>cctccagcac</td><td>ctggcccagc</td><td>600</td>

<td>gagaccgtca</td><td>cctgcaacgt</td><td>tgcccacccg</td><td>gccagcagca</td><td>ccaaggtgga</td><td>caagaaaatt</td><td>660</td>

<td>gtgcccaggg</td><td>attgtggttg</td><td>taagccttgc</td><td>atatgtacag</td><td>tcccagaagt</td><td>atcatctgtc</td><td>720</td>

<td>ttcatcttcc</td><td>ccccaaagcc</td><td>caaggatgtg</td><td>ctcaccatta</td><td>ctctgactcc</td><td>taaggtcacg</td><td>780</td>

<td>tgtgttgtgg</td><td>tagacatcag</td><td>caaggatgat</td><td>cccgaggtcc</td><td>agttcagctg</td><td>gtttgtagat</td><td>840</td>

<td>gatgtggagg</td><td>tgcacacagc</td><td>tcagacgcaa</td><td>ccccgggagg</td><td>agcagttcaa</td><td>cagcactttc</td><td>900</td>

<td>cgctcagtca</td><td>gtgaacttcc</td><td>catcatgcac</td><td>caggactggc</td><td>tcaatggcaa</td><td>ggagttcaaa</td><td>960</td>

<td>tgcagggtca</td><td>acagtgcagc</td><td>tttccctgcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>ggcagaccga</td><td>aggctccaca</td><td>ggtgtacacc</td><td>attccacctc</td><td>ccaaggagca</td><td>gatggccaag</td><td>1080</td>

<td>gataaagtca</td><td>gtctgacctg</td><td>catgataaca</td><td>gacttcttcc</td><td>ctgaagacat</td><td>tactgtggag</td><td>1140</td>

<td>tggcagtgga</td><td>atgggcagcc</td><td>agcggagaac</td><td>tacaagaaca</td><td>ctcagcccat</td><td>catggacaca</td><td>1200</td>

<td>gatggctctt</td><td>acttcatcta</td><td>cagcaagctc</td><td>aatgtgcaga</td><td>agagcaactg</td><td>ggaggcagga</td><td>1260</td>

<td>aatactttca</td><td>cctgctctgt</td><td>gttacatgag</td><td>ggcctgcaca</td><td>accaccatac</td><td>tgagaagagc</td><td>1320</td>

<td>ctctcccact</td><td>ctcctggtaa</td><td>and</td><td></td><td></td><td></td><td>1341</td>

<210> 110 <211> 447 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN. <223> Ab-7 heavy chain <400> 110

PZ / 4572 / AGR 100

EP 2 569 010 B1

<td rowspan="2">Glu 1</td><td rowspan="2">val</td><td rowspan="2">Gin</td><td colspan="9">Leu Gin Gin Cheese Gly Pro Glu Leu Met</td><td colspan="2" rowspan="2">Lys Pro</td><td rowspan="2">Gly 15</td><td rowspan="2">ala</td>

<td colspan="2">5</td><td colspan="7">10</td>

<td>Cheese</td><td>val</td><td>lys</td><td>Underworld</td><td>Cheese</td><td>Cy3</td><td>lys</td><td>ala</td><td>Cheese</td><td>Gly</td><td>Tyr</td><td>Thr</td><td>phe</td><td>Thr</td><td>Asp</td><td>Tyr</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>own</td><td>Underworld</td><td>His</td><td>Trp</td><td>Underworld</td><td>lys</td><td>Gin</td><td>own</td><td>Gin</td><td>Gly</td><td>Ly3</td><td>Cheese</td><td>Leu</td><td>Glu</td><td>Trp</td><td>How much</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Gly</td><td>Glu</td><td>How much</td><td>own</td><td>Pro</td><td>Α3Π</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>ala</td><td>Gly</td><td>Tyr</td><td>own</td><td>Gin</td><td>Gin</td><td>phe</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>lys</td><td>Gly</td><td>Ly3</td><td>ala</td><td>Thr</td><td>Leu</td><td>Thr</td><td>val</td><td>Asp</td><td>Ly3</td><td>Cheese</td><td>Cheese</td><td>Arg</td><td>Thr</td><td>ala</td><td>Tyr</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Underworld</td><td>Glu</td><td>Leu</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>A3P</td><td>Cheese</td><td>ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cys</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>ala</td><td>Arg</td><td>Leu</td><td>Gly</td><td>Tyr</td><td>val</td><td>Gly</td><td>own</td><td>Tyr</td><td>Glu</td><td>Asp</td><td>Trp</td><td>Tyr</td><td>phe</td><td>Asp</td><td>val</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>Trp</td><td>Gly</td><td>ala</td><td>Gly</td><td>Thr</td><td>Thr</td><td>val</td><td>Thr</td><td>val</td><td>Cheese</td><td>Cheese</td><td>ala</td><td>Ly3</td><td>Thr</td><td>Thr</td><td>Pro</td>

<td></td><td></td><td>115</td><td></td><td></td><td></td><td></td><td>120</td><td></td><td></td><td></td><td></td><td>125</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>val</td><td>Tyr</td><td>Pro</td><td>Leu</td><td>ala</td><td>Pro</td><td>Gly</td><td>Cheese</td><td>ala</td><td>ala</td><td>Gin</td><td>Thr</td><td>own</td><td>Cheese</td>

VP / 4572 / AGR

101

EP 2 569 010 B1

130 135 140

Met Val Thr Leu Gly Cys Leu Val Ly3 Gly Tyr Phe Pro Glu Pro Val 145 150 155 160

Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe 165 170 175

Pro Ala Val Leu Gin Cheese Asp Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr 180 185 190

Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala 195 200 205

His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg A3p 210 215 220

Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val 225 230 235 240

Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr 245 250 255

Pro Lye Val Thr Cys Val Val Val Asp. Ile Ser Lys Asp Asp Pro Glu 260 265 270

Val Gin Phe Ser Trp Phe Val A3p Asp Val Glu Val His Thr Ala Gin 275 280 285

Thr Gin Pro Arg Glu Glu Gin Phe Asn Cheese Thr Phe Arg Cheese Val Ser 290 295 300

Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Ly3 305 310 315 320

Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Ile Glu Lys Thr Ile 325 330 335

Ser Ly3 Thr Lys Gly Arg Pro Lys Al Pro Gin Val Tyr Thr Ile Pro 340 345 350

Pro Pro Lys Glu Gin Met Ala Lys A3p Lys Val Ser Leu Thr Cys Met 355 360 365

How much Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn

VP / 4572 / AGR

102

EP 2 569 010 B1

370

375

380

Gly Gin Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro How Much Met A3p Thr 385 390 395 400

Asp Gly Cheese Tyr Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys Cheese Asn 405 410 415

Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu 420 425 430

His Asn His Thr Glu Lys Ser Leu Ser His Cheese Pro Gly Lys 435 440 445 <210> 111 <211> 11 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-8 <400> 111

Arg Ala Ser Gin Asp Ile Ser Asn Tyr Leu Asn

10 <210> 112 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-8 <400> 112

Tyr Thr Ser Arg Leu Leu Cheese

5 <210> 113 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-8 <400> 113

Gin Gin Gly Asp Thr Leu Pro Tyr Thr

5 <210> 114 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-8 <400> 114

Asp Tyr Aen Met His

5 <210> 115 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-8 <400> 115

Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr Asn Gin Lys Phe Lys 15 10 15

Gly

VP / 4572 / AGR

103

EP 2 569 010 B1 <210> 116 <211> 14 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-8 <400> 116

Leu Gly Tyr Asp Asp. Ile Asp. Asp A3p Trp Tyr Phe Asp Val 15 10 <210> 117 <211> 645 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-8 light chain <400> 117

<td>gatatccaga</td><td>tgacacagac</td><td>tacatcctcc</td><td>ctgtctgcct</td><td>ctctgggaga</td><td>cagggtctcc</td><td>60</td>

<td>atcagttgca</td><td>gggcaagtca</td><td>agacattagc</td><td>aattatttaa</td><td>actggtatca</td><td>gcagaaacca</td><td>120</td>

<td>gatggaactt</td><td>ttaaactcct</td><td>tatcttctac</td><td>acatcaagat</td><td>tactctcagg</td><td>agtcccatca</td><td>180</td>

<td>aggttcagtg</td><td>gcagtgggtc</td><td>tggaacagat</td><td>tattctctca</td><td>ccatttacaa</td><td>cctggagcaa</td><td>240</td>

<td>gaagattttg</td><td>ccacttactt</td><td>ttgccaacag</td><td>ggagatacgc</td><td>ttccgtacac</td><td>tttcggaggg</td><td>300</td>

<td>gggaccaaac</td><td>tggaaataaa</td><td>acgggctgat</td><td>gctgcaccaa</td><td>ctgtatccat</td><td>cttcccacca</td><td>360</td>

<td>tccagtgagc</td><td>agttaacatc</td><td>tggaggtgcc</td><td>tcagtcgtgt</td><td>gcttcttgaa</td><td>caacttctac</td><td>420</td>

<td>cccaaagaca</td><td>tcaatgtcaa</td><td>gtggaagatt</td><td>gatggcagtg</td><td>aacgacaaaa</td><td>tggcgtcctg</td><td>480</td>

<td>aacagttgga</td><td>ctgatcagga</td><td>cagcaaagac</td><td>agcacctaca</td><td>gcatgagcag</td><td>caccctcacg</td><td>540</td>

<td>ttgaccaagg</td><td>acgagtatga</td><td>acgacataac</td><td>agctatacct</td><td>gtgaggccac</td><td>tcacaagaca</td><td>600</td>

<td>tcaacttcac</td><td>ccattgtcaa</td><td>gagcttcaac</td><td>aggaatgagt</td><td>gttag</td><td></td><td>645</td>

<210> 118 <211> 214 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-8 light chain <400> 118

VP / 4572 / AGR

104

EP 2 569 010 B1

Asp Ile Gin Met Thr Gin Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 15 10 15

Asp Arg. Val Ser. Ser. Ser Cys. Arg. Ala. Ser Gin. Asp. Ser Asn. Tyr 20 25 30

Leu Asn Trp Tyr Gin Gin Lys Pro Asp Gly Thr Phe Lys Leu Leu Ile 35 40 45

Phe Tyr Thr Ser Arg Leu Leu Cheese Gly Val Pro Cheese Arg Phe Cheese Gly 50 55 60

Cheese Gly Cheese Gly Thr Asp Tyr Cheese Leu Thr Ile Tyr Asn Leu Glu Gin 65 70 75 80

Glu Asp Phe Ala Thr Tyr Phe Cys Gin Gin Gly Asp Thr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110

Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gin Leu Thr Cheese Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140

Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu 145 150 155 160

A3n Ser Trp Thr Asp Gin Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Cheese Tyr 180 185 190

Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Ly3 Ser 195 200 205

Phe Asn Arg Asn Glu Cys 210 <210> 119 <211> 1344 <212> DNA <213> Mus musculus <220>

<221> trait_different

<223> Ab-8 heavy chain <400> 119

VP / 4572 / AGR

105

EP 2 569 010 B1

<td>gaggtccaac</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctaatgaagc</td><td>ctggggcttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggata</td><td>tacattcact</td><td>gactacaaca</td><td>tgcactgggt</td><td>gaagcagaac</td><td>120</td>

<td>caaggaaaga</td><td>ccctagactg</td><td>gataggagaa</td><td>attaatccta</td><td>acagtggtgg</td><td>tgctggctac</td><td>180</td>

<td>aaccagaagt</td><td>tcaagggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcctccac</td><td>cacagcctac</td><td>240</td>

<td>atggagctcc</td><td>gcagcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagattgggc</td><td>300</td>

<td>tacgatgata</td><td>tctacgacga</td><td>ctggtacttc</td><td>gatgtctggg</td><td>gcgcagggac</td><td>cacggtcacc</td><td>360</td>

<td>gtctcctcag</td><td>ccaaaacgac</td><td>acccccatct</td><td>gtctatccac</td><td>tggcccctgg</td><td>atctgctgcc</td><td>420</td>

<td>caaactaact</td><td>ccatggtgac</td><td>cctgggatgc</td><td>ctggtcaagg</td><td>gctatttccc</td><td>tgagccagtg</td><td>480</td>

<td>acagtgacct</td><td>ggaactctgg</td><td>atccctgtcc</td><td>agcggtgtgc</td><td>acaccttccc</td><td>agctgtcctg</td><td>540</td>

<td>cagtctgacc</td><td>tctacactct</td><td>gagcagctca</td><td>gtgactgtcc</td><td>cctccagcac</td><td>ctggcccagc</td><td>600</td>

<td>gagaccgtca</td><td>cctgcaacgt</td><td>tgcccacccg</td><td>gccagcagca</td><td>ccaaggtgga</td><td>caagaaaatt</td><td>660</td>

<td>gtgcccaggg</td><td>attgtggttg</td><td>taagccttgc</td><td>atatgtacag</td><td>tcccagaagt</td><td>atcatctgtc</td><td>720</td>

<td>ttcatcttcc</td><td>ccccaaagcc</td><td>caaggatgtg</td><td>ctcaccatta</td><td>ctctgactcc</td><td>taaggtcacg</td><td>780</td>

<td>tgtgttgtgg</td><td>tagacatcag</td><td>caaggatgat</td><td>cccgaggtcc</td><td>agttcagctg</td><td>gtttgtagat</td><td>840</td>

<td>gatgtggagg</td><td>tgcacacagc</td><td>tcagacgcaa</td><td>ccccgggagg</td><td>agcagttcaa</td><td>cagcactttc</td><td>900</td>

<td>cgctcagtca</td><td>gtgaacttcc</td><td>catcatgcac</td><td>caggactggc</td><td>tcaatggcaa</td><td>ggagttcaaa</td><td>960</td>

<td>tgcagggtca</td><td>acagtgcagc</td><td>tttccctgcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>ggcagaccga</td><td>aggctccaca</td><td>ggtgtacacc</td><td>attccacctc</td><td>ccaaggagca</td><td>gatggccaag</td><td>1080</td>

<td>gataaagtca</td><td>gtctgacctg</td><td>catgataaca</td><td>gacttcttcc</td><td>ctgaagacat</td><td>tactgtggag</td><td>1140</td>

<td>tggcagtgga</td><td>atgggcagcc</td><td>agcggagaac</td><td>tacaagaaca</td><td>ctcagcccat</td><td>catggacaca</td><td>1200</td>

<td>gatggctctt</td><td>acttcatcta</td><td>cagcaagctc</td><td>aatgtgcaga</td><td>agagcaactg</td><td>ggaggcagga</td><td>1260</td>

<td>aatactttca</td><td>cctgctctgt</td><td>gttacatgag</td><td>ggcctgcaca</td><td>accaccatac</td><td>tgagaagagc</td><td>1320</td>

<td>ctctcccact</td><td>ctcctggtaa</td><td>atga</td><td></td><td></td><td></td><td>1344</td>

<210> 120 <211> 447 <212> PRT <213> Mus musculus <220>

<221> CECHA_RÓŻN.

<223> Ab-8 heavy chain <400> 120

EP 2 569 010 B1

VP / 4572 / AGR

Glu Val Gin Leu Gin Gin Cheese Gly 1 5

106

Pro Glu Leu Met Lys Pro Gly Ala 10 15

Ser Val Lye Met Ser Cys Lys Ala 20

Gly Tyr Thr Phe Thr Asp Tyr 25 30

Asn Met His Trp Val Lys Gin Asn 35 40

Gin Gly Lys Thr Leu Asp Trp Ile 45

Gly Glu Ile Asn Pro Asn Cheese Gly 50 55

Gly Ala Gly Tyr Asn Gin Lys Phe 60

Lys Gly Lys Ala Thr Leu Thr Val 65 TO

Asp Lys Ser Ser Thr Thr Ala Tyr 75 80

Met Glu Leu Arg Ser Leu Thr Ser 85

Glu Asp Ser Ala Val Tyr Tyr Cys 90 95

Ala Arg Leu Gly Tyr Asp Asp Ile

Tyr Asp Asp Trp Tyr Phe Asp Val

VP / 4572 / AGR

107

EP 2 569 010 B1

100 105 110

Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Al Lys Thr Thr Pro 115 120 125

Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr Asn Cheese 130 135 140

Met Val Thr Leu Gly Cy3 Leu Val Lys Gly Tyr Phe Pro Glu Pro Val 145 150 155 160

Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe 165 170 175

Pro Ala Val Leu Gin Cheese Asp Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr 180 185 190

Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala 195 200 205

His Pro Ala Ser Ser Thr Ly3 Val Asp Lys Lys Ile Val Pro Arg Asp 210 215 220

Cys Gly Cy3 Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val 225 230 235 240

Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr 245 250 255

Pro Lys Val Thr Cys Val Val Val Asp. Ile Ser Lys Asp Asp Pro Glu 260 265 270

Val Gin Phe Ser Trp Phe Val Asp Asp. Val Gl. Val. His Thr. Ala Gin 275 280 285

Thr Gin Pro Arg Glu Glu Gin Phe Asn Cheese Thr Phe Arg Cheese Val Ser 290 295 300

Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Lys 305 310 315 320

Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Ile Glu Lys Thr Ile 325 330 335

Lys Thr Lys Gly Arg Pro Lys Al Pro Gin Val Tyr Thr Ile Pro

VP / 4572 / AGR

108

EP 2 569 010 B1

340

345

350

Pro Pro Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met 355 360 365

How much Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn 370 375 380

Gly Gin Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro Ile Met Asp Thr 385 390 395 400

Asp Gly Cheese Tyr Phe Ile Tyr Cheese Lys Leu Aen Val Gin Lys Cheese Asn 405 410 415

Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu 420 425 430

His Asn His Thr Glu Lys Ser Leu Ser His Cheese Pro Gly Lys 435 440 445 <210> 121 < 211> 11 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-9 <400> 121

Arg Ala Ser Gin Asp Ile Ser Asn Tyr Leu Asn

10 <210> 122 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-9 <400> 122

Tyr Thr Ser Arg Leu Phe Ser

5 <210> 123 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-9 <400> 123

Gin Gin Gly Asp Thr Leu Pro Tyr Thr

5 <210> 124 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-9 <400> 124

Asp Tyr Asn Met His

5 <210> 125 <211> 17 <212> PRT <213> Artificial sequence

VP / 4572 / AGR

109

EP 2 569 010 B1 <220>

<223> HCDR2 Ab-9 <400> 125

Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr Asn Gin Lys Phe Lys 15 10 15

Gly <210> 126 <211> 14 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-9 <400> 126

Leu Gly Tyr Asp Asp. Ile Asp. Asp Asp Trp Tyr Phe Asp Val 1 5 10 <210> 127 <211> 642 <212> DNA <213> Mus musculus <400> 127

<td>gatatccaga</td><td>tgacacagat</td><td>tacatcctcc</td><td>ctgtctgcct</td><td>ctctgggaga</td><td>cagggtctcc</td><td>60</td>

<td>atcagttgca</td><td>gggcaagtca</td><td>agacattagc</td><td>aattatttaa</td><td>attggtatca</td><td>gcagaaacca</td><td>120</td>

<td>gatggaactt</td><td>ttaaactcct</td><td>tatcttctac</td><td>acatcaagat</td><td>tattttcagg</td><td>agtcccatca</td><td>180</td>

<td>aggttcagtg</td><td>gcagtgggtc</td><td>tggaacagat</td><td>tattctctca</td><td>ccatttacaa</td><td>cctggagcaa</td><td>240</td>

<td>gaagattttg</td><td>ccacttactt</td><td>ttgccaacag</td><td>ggagatacgc</td><td>ttccgtacac</td><td>tttcggaggg</td><td>300</td>

<td>gggaccaagg</td><td>tggaaataaa</td><td>acgggctgat</td><td>gctgcaccaa</td><td>ctgtatccat</td><td>cttcccacca</td><td>360</td>

<td>tccagtgagc</td><td>agttaacatc</td><td>tggaggtgcc</td><td>tcagtcgtgt</td><td>gcttcttgaa</td><td>caacttctac</td><td>420</td>

<td>cccaaagaca</td><td>tcaatgtcaa</td><td>gtggaagatt</td><td>gatggcagtg</td><td>aacgacaaaa</td><td>tggcgtcctg</td><td>480</td>

<td>aacagttgga</td><td>ctgatcagga</td><td>cagcaaagac</td><td>agcacctaca</td><td>gcatgagcag</td><td>caccctcacg</td><td>540</td>

<td>ttgaccaagg</td><td>acgagtatga</td><td>acgacataac</td><td>agctatacct</td><td>gtgaggccac</td><td>tcacaagaca</td><td>600</td>

<td>tcaacttcac</td><td>ccattgtcaa</td><td>gagcttcaac</td><td>aggaatgagt</td><td>gt</td><td></td><td>642</td>

<210> 128 <211> 214 <212> PRT <213> Mus musculus <400> 128

VP / 4572 / AGR

110

EP 2 569 010 B1

<td>Asp 1</td><td colspan="3">Ile Gin Met</td><td>Thr 5</td><td>Gin</td><td>How much</td><td>Thr</td><td>Cheese</td><td>Cheese 10</td><td>Leu</td><td>Cheese</td><td>ala</td><td>Cheese</td><td>Leu 15</td><td>Gly</td>

<td>Asp</td><td>Arg</td><td>val</td><td>Cheese</td><td>How much</td><td>Cheese</td><td>Cys</td><td>Arg</td><td>ala</td><td>Cheese</td><td>Gin</td><td>Asp</td><td>How much</td><td>Cheese</td><td>own</td><td>Tyr</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>Leu</td><td>own</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>lys</td><td>Pro</td><td>Asp</td><td>Gly</td><td>Thr</td><td>phe</td><td>lys</td><td>Leu</td><td>Leu</td><td>How much</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>phe</td><td>Tyr</td><td>Thr</td><td>Cheese</td><td>Arg</td><td>Leu</td><td>phe</td><td>Cheese</td><td>Gly</td><td>val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>phe</td><td>Cheese</td><td>Gly</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Cheese</td><td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Asp</td><td>Tyr</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>How much</td><td>Tyr</td><td>own</td><td>Leu</td><td>Glu</td><td>Gin</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Glu</td><td>Asp</td><td>phe</td><td>ala</td><td>Thr</td><td>Tyr</td><td>phe</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Gly</td><td>Asp</td><td>Thr</td><td>Leu</td><td>Pro</td><td>Tyr</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>Thr</td><td>phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>lys</td><td>val</td><td>Glu</td><td>How much</td><td>lys</td><td>Arg</td><td>ala</td><td>Asp</td><td>ala</td><td>ala</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>Pro</td><td>Thr</td><td>val</td><td>Cheese</td><td>How much</td><td>phe</td><td>Pro</td><td>Pro</td><td>Cheese</td><td>Cheese</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Gly</td>

<td></td><td></td><td>115</td><td></td><td></td><td></td><td></td><td>120</td><td></td><td></td><td></td><td></td><td>125</td><td></td><td></td><td></td>

<td>Gly</td><td>ala</td><td>Cheese</td><td>val</td><td>val</td><td>Cys</td><td>phe</td><td>Leu</td><td>own</td><td>own</td><td>phe</td><td>Tyr</td><td>Pro</td><td>lys</td><td>Asp</td><td>How much</td>

<td></td><td>130</td><td></td><td></td><td></td><td></td><td>135</td><td></td><td></td><td></td><td></td><td>140</td><td></td><td></td><td></td><td></td>

<td>own</td><td>val</td><td>lys</td><td>Trp</td><td>lys</td><td>How much</td><td>Asp</td><td>Gly</td><td>Cheese</td><td>Glu</td><td>Arg</td><td>Gin</td><td>own</td><td>Gly</td><td>val</td><td>Leu</td>

<td>145</td><td></td><td></td><td></td><td></td><td>150</td><td></td><td></td><td></td><td></td><td>155</td><td></td><td></td><td></td><td></td><td>160</td>

<td>own</td><td>Cheese</td><td>Trp</td><td>Thr</td><td>A3P</td><td>Gin</td><td>Asp</td><td>Cheese</td><td>lys</td><td>Asp</td><td>Cheese</td><td>Thr</td><td>Tyr</td><td>Cheese</td><td>Underworld</td><td>Cheese</td>

<td></td><td></td><td></td><td></td><td>165</td><td></td><td></td><td></td><td></td><td>170</td><td></td><td></td><td></td><td></td><td>175</td><td></td>

<td>Cheese</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Thr</td><td>lys</td><td>Asp</td><td>Glu</td><td>Tyr</td><td>Glu</td><td>Arg</td><td>His</td><td>own</td><td>Cheese</td><td>Tyr</td>

<td></td><td></td><td></td><td>180</td><td></td><td></td><td></td><td></td><td>185</td><td></td><td></td><td></td><td></td><td>190</td><td></td><td></td>

<td>Thr</td><td>Cys</td><td>Glu</td><td>ala</td><td>Thr</td><td>His</td><td>lys</td><td>Thr</td><td>Cheese</td><td>Thr</td><td>Cheese</td><td>Pro</td><td>How much</td><td>val</td><td>lys</td><td>Cheese</td>

<td></td><td></td><td>195</td><td></td><td></td><td></td><td></td><td>200</td><td></td><td></td><td></td><td></td><td>205</td><td></td><td></td><td></td>

<td>phe</td><td>own</td><td>Arg</td><td>own</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

210 <210> 129 <211> 1350 <212> DNA <213> Mus musculus <400> 129

VP / 4572 / AGR

111

EP 2 569 010 B1

<td>gaggtccaac</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctaatgaagc</td><td>ctgggacttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggata</td><td>tacattcact</td><td>gactacaaca</td><td>tgcactgggt</td><td>gaagcagacc</td><td>120</td>

<td>caaggaaaga</td><td>ccctagagtg</td><td>gataggagaa</td><td>attaatccta</td><td>acagtggtgg</td><td>tgctggctac</td><td>180</td>

<td>aaccagaagt</td><td>tcaagggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcctccac</td><td>cacagcctac</td><td>240</td>

<td>atggagctcc</td><td>gcagcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aaaattgggc</td><td>300</td>

<td>tacgatgata</td><td>tctacgacga</td><td>ctggtatttc</td><td>gatgtctggg</td><td>gcgcagggac</td><td>cacggtcacc</td><td>360</td>

<td>gtctcctcag</td><td>ccaaaacaac</td><td>agccccatcg</td><td>gtctatccac</td><td>tggcccctgt</td><td>gtgtggagat</td><td>420</td>

<td>acaactggct</td><td>cctcggtgac</td><td>tctaggatgc</td><td>ctggtcaagg</td><td>gttatttccc</td><td>tgagccagtg</td><td>480</td>

<td>accttgacct</td><td>ggaactctgg</td><td>atccctgtcc</td><td>agtgatgtgc</td><td>acaccttccc</td><td>agctctcctg</td><td>540</td>

<td>cagtctggcc</td><td>tctacaccct</td><td>cagcagctca</td><td>gtgactgtaa</td><td>ccacctggcc</td><td>cagccagacc</td><td>600</td>

<td>atcacctgca</td><td>atgtggccca</td><td>cccggcaagc</td><td>agcaccaaag</td><td>tggacaagaa</td><td>aattgagccc</td><td>660</td>

<td>agagggtccc</td><td>caacacataa</td><td>accctgtcct</td><td>ccatgcccag</td><td>ctcctaacct</td><td>cttgggtgga</td><td>720</td>

<td>ccatccgtct</td><td>tcatcttccc</td><td>tccaaagatc</td><td>aaggatgtac</td><td>tcatgatctc</td><td>cctgagcccc</td><td>780</td>

<td>atggtcacgt</td><td>gtgtggtggt</td><td>ggatgtgagc</td><td>gaggatgacc</td><td>cagatgtcca</td><td>tgtcagctgg</td><td>840</td>

<td>ttcgtgaaca</td><td>acgtggaagt</td><td>acacacagct</td><td>cagacacaaa</td><td>cccatagaga</td><td>ggattacaac</td><td>900</td>

<td>agtactatcc</td><td>gggtggtcag</td><td>tgccctcccc</td><td>atccagcacc</td><td>aggactggat</td><td>gagtggcaag</td><td>960</td>

<td>gagttcaaat</td><td>gcaaggtcaa</td><td>caacaaagcc</td><td>ctcccagcgc</td><td>ccatcgagag</td><td>aaccatctca</td><td>1020</td>

<td>aaacccaaag</td><td>ggccagtaag</td><td>agctccacag</td><td>gtatatgtct</td><td>tgcctccacc</td><td>agaagaagag</td><td>1080</td>

<td>atgactaaga</td><td>aacaggtcac</td><td>tctgacctgc</td><td>atgatcacag</td><td>acttcatgcc</td><td>tgaagacatt</td><td>1140</td>

<td>tacgtggagt</td><td>ggaccaacaa</td><td>cgggcaaaca</td><td>gagctaaact</td><td>acaagaacac</td><td>tgaaccagtc</td><td>1200</td>

<td>ctggactctg</td><td>atggttctta</td><td>cttcatgtac</td><td>agcaagctga</td><td>gagtggaaaa</td><td>gaagaactgg</td><td>1260</td>

<td>gtggaaagaa</td><td>atagctactc</td><td>ctgttcagtg</td><td>gtccacgagg</td><td>gtctgcacaa</td><td>tcaccacacg</td><td>1320</td>

<td>actaagagct</td><td>tctcccggac</td><td>tccgggtaaa</td><td></td><td></td><td></td><td>1350</td>

<210> 130 <211> 450 <212> PRT <213> Mus musculus <400> 130

VP / 4572 / AGR

112

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Cheese Gly Pro Glu Leu Met Lys Pro Gly Thr 15 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

A3n Met His Trp Val Lys Gin Thr Gin Gly Lys Thr Leu Glu Trp Ile 35 40 45

Gly Glu Ile Asn Pro A3n Ser Gly Gly Ala Gly Tyr Asn Gin Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Thr Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Leu Gly Tyr Asp Asp. Ile Tyr A3p Asp Trp Tyr Phe Asp Val 100 105 110

Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Ala Lys Thr Thr Ala 115 120 125

Pro Ser Val Tyr Pro Leu Ala Pro Val Cys Gly Asp Thr Thr Gly Cheese 130 135 140

Cheese Val Thr Leu Gly Cy3 Leu Val Lys Gly Tyr Phe Pro Glu Pro Val

VP / 4572 / AGR

113

EP 2 569 010 B1

145 150 155 160

Thr Leu Thr Trp Asn Cheese Gly Cheese Leu Cheese Cheese Asp Val His Thr Phe 165 170 175

Pro Ala Leu Leu Gin Cheese Gly Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr 180 185 190

Val Thr Thr Trp Pro Ser Gin Thr Ile Thr Cys Asn Val Ala His Pro 195 200 205

Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Glu Pro Arg Gly Pro Pro 210 215 220

Thr His Lys Pro Cys Pro Pro Cys Pro Al Pro A3n Leu Leu Gly Gly 225 230 235 240

Pro Ser Val Phe Ile Phe Pro Pro Ly3 Ile Lys Asp Val Leu Met Ile 245 250 255

Ser Leu Ser Pro Met Val Thr Cys Val Val Val Asp. Val Ser Glu Asp 260 265 270

Asp Pro Asp Val Hi3 Val Ser Trp Phe Val Asn Asn Val Glu Val His 275 280 285

Thr Ala Gin Thr Gin Thr His Arg Glu Asp Tyr Asn Ser Thr Ile Arg 290 295 300

Val Val Ser Ala Leu Pro Ile Gin His Gin Asp Trp Met Ser Gly Lys 305 310 315 320

Glu Phe Lys Cys Lys Val Asn A3n Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335

Arg Thr Ile Ly3 Pro Pro Lys Gly Pro Val Arg Ala Pro Gin Val Tyr 340 345 350

Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys Ly3 Gin Val Thr Leu 355 360 365

Thr Cys Met Ile Thr Asp Phe Met Pro Glu Asp Ile Tyr Val Glu Trp 370 375 380

Thr A3n Asn Gly Gin Thr Glu Leu Asn Tyr Lys Asn Thr Glu Pro Val

VP / 4572 / AGR

114

EP 2 569 010 B1

390

395

400

Leu Asp Ser Asp Gly Cheese Tyr Phe Met Tyr Cheese Lys Leu Arg Val Glu 405 410 415

Lys Lys Asn Trp Val Glu Arg Aen Ser Tyr Ser Cys Ser Val Val His 420 425 430

Glu Gly Leu His Asn Hi3 His Thr Thr Lys Ser Phe Ser Arg Thr Pro 435 440 445

Gly Lys 450 <210> 131 <211> 11 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-10 <400> 131

Arg Ala Ser Gin Asp Ile Ser Asn Tyr Leu Asn

10 <210> 132 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-10 <400> 132

Tyr Thr Ser Arg Leu Leu Cheese

5 <210> 133 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-10 <400> 133

Gin Gin Gly Asp Thr Leu Pro Tyr Thr

5 <210> 134 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-10 <400> 134

Asp Tyr Asn Met His

5 <210> 135 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-10 <400> 135

Glu Ile Asn Pro Asn Ser Gly Gly Ala Gly Tyr Asn Gin Lys Phe Lys 15 10 15

Gly

VP / 4572 / AGR

115

EP 2 569 010 B1 <210> 136 <211> 14 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-10 <400> 136

Leu Gly Tyr Asp Asp Lyr Asp Asp Trp Tyr Phe Asp Val

5 10 <210> 137 <211> 645 <212> DNA <213> Mus musculus <400> 137 gatatccaga tgacacagac tacatcctcc ctgtctgcct ctctgggaga cagggtctcc 60 atcagttgca gggcaagtca agacattagc aattatttaa actggtatca gcagaaacca 120 gatggaactt ttaaactcct tatcttctac acatcaagat tactctcagg agtcccatca 180 aggttcagtg gcagtgggtc tggaacagat tattctctca ccatttacaa cctggagcaa 240 gaagattttg ccacttactt ttgccaacag ggagatacgc ttccgtacac tttcggaggg 300 gggaccaaac tggaaataaa acgggctgat gctgcaccaa ctgtatccat cttcccacta 360 tccagtgagc agttaacatc tggaggtgcc tcagtcgtgt gcttcttgaa caacttctac 420 cccaaagaca tcaatgtcaa gtggaagatt gatggcagtg aacgacaaaa tggcgtcctg 480 aacagttgga ctgatcagga cagcaaagac agcacctaca gcatgagcag caccctcacg 540 ttgaccaagg acgagtatga acgacataac agctatacct gtgaggccac tcacaagaca 600 tcaacttcac ccattgtcaa gagcttcaac aggaatgagt gttag 645 < 210>138 <211> 214 <212> PRT <213> Mus musculus <400> 138

VP / 4572 / AGR

116

EP 2 569 010 B1

Asp Ile Gin Met Thr Gin Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 15 10 15

Asp Arg. Val Ser. Ser. Ser Cys. Arg. Ala. Ser Gin. Asp. Ser Asn. Tyr 20 25 30

Leu Asn Trp Tyr Gin Gin Lys Pro Asp Gly Thr Phe Lys Leu Leu Ile 35 40 45

Phe Tyr Thr Ser Arg Leu Leu Cheese Gly Val Pro Cheese Arg Phe Cheese Gly 50 55 60

Cheese Gly Cheese Gly Thr Asp Tyr Cheese Leu Thr Ile Tyr Asn Leu Glu Gin 65 70 75 80

Glu Asp Phe Ala Thr Tyr Phe Cys Gin Gin Gly Asp Thr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110

Pro Thr Val Ser Ile Phe Pro Leu Cheese Ser Glu Gin Leu Thr Cheese Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn A3n Phe Tyr Pro Lys Asp Ile 130 135 140

Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu 145 150 155 160

Asn Ser Trp Thr Asp Gin Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Cheese Tyr 180 185 190

Thr Cys Glu Ala Thr Hi3 Lys Thr Ser Thr Ser Pro Ile Val Lye Ser 195 200 205

Phe Asn Arg Asn Glu Cy3 210 <210> 139 <211> 1344 <212> DNA <213> Mus musculus <400> 139

VP / 4572 / AGR

117

EP 2 569 010 B1

<td>gaggtccaac</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctaatgaagc</td><td>ctggggcttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggata</td><td>tacattcact</td><td>gactacaaca</td><td>tgcactgggt</td><td>gaagcagaac</td><td>120</td>

<td>caaggaaaga</td><td>ccctagaatg</td><td>gataggagaa</td><td>attaatccta</td><td>acagtggtgg</td><td>tgctggctac</td><td>180</td>

<td>aaccagaagt</td><td>tcaagggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcctccac</td><td>cacagcctac</td><td>240</td>

<td>atggagctcc</td><td>gcagcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagattgggc</td><td>300</td>

<td>tacgatgata</td><td>tctacgacga</td><td>ctggtacttc</td><td>gatgtctggg</td><td>gcgcagggac</td><td>cacggtcacc</td><td>360</td>

<td>gtctcctcag</td><td>ccaaaacgac</td><td>acccccatct</td><td>gtctatccac</td><td>tggcccctgg</td><td>atctgctgcc</td><td>420</td>

<td>caaactaact</td><td>ccatggtgac</td><td>cctgggatgc</td><td>ctggtcaagg</td><td>gctatttccc</td><td>tgagccagtg</td><td>480</td>

<td>acagtgacct</td><td>ggaactctgg</td><td>atccctgtcc</td><td>agcggtgtgc</td><td>acaccttccc</td><td>agctgtcctg</td><td>540</td>

<td>cagtctgacc</td><td>tctacactct</td><td>gagcagctca</td><td>gtgactgtcc</td><td>cctccagcac</td><td>ctggcccagc</td><td>600</td>

<td>gagaccgtca</td><td>cctgcaacgt</td><td>tgcccacccg</td><td>gccagcagca</td><td>ccaaggtgga</td><td>caagaaaatt</td><td>660</td>

<td>gtgcccaggg</td><td>attgtggttg</td><td>taagccttgc</td><td>atatgtacag</td><td>tcccagaagt</td><td>atcatctgtc</td><td>720</td>

<td>ttcatcttcc</td><td>ccccaaagcc</td><td>caaggatgtg</td><td>ctcaccatta</td><td>ctctgactcc</td><td>taaggtcacg</td><td>780</td>

<td>tgtgttgtgg</td><td>tagacatcag</td><td>caaggatgat</td><td>cccgaggtcc</td><td>agttcagctg</td><td>gtttgtagat</td><td>840</td>

<td>gatgtggagg</td><td>tgcacacagc</td><td>tcagacgcaa</td><td>ccccgggagg</td><td>agcagttcaa</td><td>cagcactttc</td><td>900</td>

<td>cgctcagtca</td><td>gtgaacttcc</td><td>catcatgcac</td><td>caggactggc</td><td>tcaatggcaa</td><td>ggagttcaaa</td><td>960</td>

<td>tgcagggtca</td><td>acagtgcagc</td><td>tttccctgcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>ggcagaccga</td><td>aggctccaca</td><td>ggtgtacacc</td><td>attccacctc</td><td>ccaaggagca</td><td>gatggccaag</td><td>1080</td>

<td>gataaagtca</td><td>gtctgacctg</td><td>catgataaca</td><td>gacttcttcc</td><td>ctgaagacat</td><td>tactgtggag</td><td>1140</td>

<td>tggcagtgga</td><td>atgggcagcc</td><td>agcggagaac</td><td>tacaagaaca</td><td>ctcagcccat</td><td>catggacaca</td><td>1200</td>

<td>gatggctctt</td><td>acttcatcta</td><td>cagcaagctc</td><td>aatgtgcaga</td><td>agagcaactg</td><td>ggaggcagga</td><td>1260</td>

<td>aatactttca</td><td>cctgctctgt</td><td>gttacatgag</td><td>ggcctgcaca</td><td>accaccatac</td><td>tgagaagagc</td><td>1320</td>

<td>ctctcccact</td><td>ctcctggtaa</td><td>atga</td><td></td><td></td><td></td><td>1344</td>

<210> 140 <211> 447 <212> PRT <213> Mus musculus <400> 140

EP 2 569 010 B1

<td colspan="9">VP / 4572 / AGR</td><td colspan="7">118</td>

<td>Glu</td><td>val</td><td>Gin</td><td>Leu</td><td>Gin</td><td>Gin</td><td>Cheese</td><td>Gly</td><td>Pro</td><td>Glu</td><td>Leu</td><td>Underworld</td><td>lys</td><td>Pro</td><td>Gly</td><td>ala</td>

<td>1</td><td></td><td></td><td></td><td>5</td><td></td><td></td><td></td><td></td><td>10</td><td></td><td></td><td></td><td></td><td>15</td><td></td>

<td>Cheese</td><td>val</td><td>lys</td><td>Underworld</td><td>Cheese</td><td>Cys</td><td>lys</td><td>ala</td><td>Cheese</td><td>Gly</td><td>Tyr</td><td>Thr</td><td>phe</td><td>Thr</td><td>Asp</td><td>Tyr</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>own</td><td>Underworld</td><td>His</td><td>Trp</td><td>val</td><td>lys</td><td>Gin</td><td>own</td><td>Gin</td><td>Gly</td><td>lys</td><td>Thr</td><td>Leu</td><td>Glu</td><td>Trp</td><td>How much</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Gly</td><td>Glu</td><td>how much</td><td>own</td><td>Pro</td><td>own</td><td>Cheese</td><td>Gly</td><td>Gly</td><td>ala</td><td>Gly</td><td>Tyr</td><td>own</td><td>Gin</td><td>lys</td><td>phe</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>lys</td><td>Gly</td><td>lys</td><td>ala</td><td>Thr</td><td>Leu</td><td>Thr</td><td>val</td><td>Asp</td><td>lys</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>Thr</td><td>ala</td><td>Tyr</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Underworld</td><td>Glu</td><td>Leu</td><td>Arg</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cheese</td><td>Glu</td><td>Asp</td><td>Cheese</td><td>ala</td><td>val</td><td>Tyr</td><td>Tyr</td><td>Cys</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>ala</td><td>Arg</td><td>Leu</td><td>Gly</td><td>Tyr</td><td>Asp</td><td>Asp</td><td>How much</td><td>Tyr</td><td>Asp</td><td>Asp</td><td>Trp</td><td>Tyr</td><td>phe</td><td>Asp</td><td>val</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>Trp</td><td>Gly</td><td>ala</td><td>Gly</td><td>Thr</td><td>Thr</td><td>val</td><td>Thr</td><td>val</td><td>Cheese</td><td>Cheese</td><td>ala</td><td>lys</td><td>Thr</td><td>Thr</td><td>Pro</td>

<td></td><td></td><td>115</td><td></td><td></td><td></td><td></td><td>120</td><td></td><td></td><td></td><td></td><td>125</td><td></td><td></td><td></td>

<td>Pro</td><td>Cheese</td><td>val</td><td>Tyr</td><td>Pro</td><td>Leu</td><td>ala</td><td>Pro</td><td>Gly</td><td>Cheese</td><td>ala</td><td>ala</td><td>Gin</td><td>Thr</td><td>own</td><td>Cheese</td>

<td></td><td>130</td><td></td><td></td><td></td><td></td><td>135</td><td></td><td></td><td></td><td></td><td>140</td><td></td><td></td><td></td><td></td>

<td>Underworld</td><td>val</td><td>Thr</td><td>Leu</td><td>Gly</td><td>Cys</td><td>Leu</td><td>val</td><td>lys</td><td>Gly</td><td>Tyr</td><td>phe</td><td>Pro</td><td>Glu</td><td>Pro</td><td>val</td>

<td>145</td><td></td><td></td><td></td><td></td><td>150</td><td></td><td></td><td></td><td></td><td>155</td><td></td><td></td><td></td><td></td><td>160</td>

<td>Thr</td><td>val</td><td>Thr</td><td>Trp</td><td>own</td><td>Cheese</td><td>Gly</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Gly</td><td>val</td><td>His</td><td>Thr</td><td>phe</td>

<td></td><td></td><td></td><td></td><td>165</td><td></td><td></td><td></td><td></td><td>170</td><td></td><td></td><td></td><td></td><td>175</td><td></td>

<td>Pro</td><td>ala</td><td>val</td><td>Leu</td><td>Gin</td><td>Cheese</td><td>Asp</td><td>Leu</td><td>Tyr</td><td>Thr</td><td>Leu</td><td>Cheese</td><td>Cheese</td><td>Cheese</td><td>val</td><td>Thr</td>

VP / 4572 / AGR

119

EP 2 569 010 B1

180 185 190

<td>val</td><td colspan="2">Pro Ser 195</td><td colspan="2">Ser Thr</td><td>Trp</td><td colspan="2">Pro Ser 200</td><td colspan="2">Glu Thr</td><td>val</td><td>Thr</td><td>Cys 205</td><td>own</td><td>val</td><td>ala</td>

<td>His</td><td>Pro</td><td>ala</td><td>Cheese</td><td>Cheese</td><td>Thr</td><td>lys</td><td>val</td><td>Asp</td><td>lys</td><td>lys</td><td>How much</td><td>val</td><td>Pro</td><td>Arg</td><td>Asp</td>

<td></td><td>210</td><td></td><td></td><td></td><td></td><td>215</td><td></td><td></td><td></td><td></td><td>220</td><td></td><td></td><td></td><td></td>

<td>Cys</td><td>Gly</td><td>Cys</td><td>lys</td><td>Pro</td><td>Cys</td><td>How much</td><td>Cy3</td><td>Thr</td><td>val</td><td>Pro</td><td>Glu</td><td>val</td><td>Cheese</td><td>Cheese</td><td>val</td>

<td>225</td><td></td><td></td><td></td><td></td><td>230</td><td></td><td></td><td></td><td></td><td>235</td><td></td><td></td><td></td><td></td><td>240</td>

<td>phe</td><td>How much</td><td>phe</td><td>Pro</td><td>Pro</td><td>lys</td><td>Pro</td><td>lys</td><td>Asp</td><td>val</td><td>Leu</td><td>Thr</td><td>How much</td><td>Thr</td><td>Leu</td><td>Thr</td>

<td></td><td></td><td></td><td></td><td>245</td><td></td><td></td><td></td><td></td><td>250</td><td></td><td></td><td></td><td></td><td>255</td><td></td>

<td>Pro</td><td>lys</td><td>val</td><td>Thr</td><td>Cys</td><td>val</td><td>val</td><td>val</td><td>Asp</td><td>How much</td><td>Cheese</td><td>lys</td><td>Asp</td><td>Asp</td><td>Pro</td><td>Glu</td>

<td></td><td></td><td></td><td>260</td><td></td><td></td><td></td><td></td><td>265</td><td></td><td></td><td></td><td></td><td>270</td><td></td><td></td>

<td>val</td><td>Gin</td><td>phe</td><td>Cheese</td><td>Trp</td><td>phe</td><td>val</td><td>Asp</td><td>Asp</td><td>val</td><td>Glu</td><td>val</td><td>His</td><td>Thr</td><td>ala</td><td>Gin</td>

<td></td><td></td><td>275</td><td></td><td></td><td></td><td></td><td>280</td><td></td><td></td><td></td><td></td><td>285</td><td></td><td></td><td></td>

<td>Thr</td><td>Gin</td><td>Pro</td><td>Arg</td><td>Glu</td><td>Glu</td><td>Gin</td><td>phe</td><td>own</td><td>Cheese</td><td>Thr</td><td>phe</td><td>Arg</td><td>Cheese</td><td>val</td><td>Cheese</td>

<td></td><td>290</td><td></td><td></td><td></td><td></td><td>295</td><td></td><td></td><td></td><td></td><td>300</td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Leu</td><td>Pro</td><td>How much</td><td>Underworld</td><td>His</td><td>Gin</td><td>A3P</td><td>Trp</td><td>Leu</td><td>own</td><td>Gly</td><td>lys</td><td>Glu</td><td>phe</td><td>Ly3</td>

<td>305</td><td></td><td></td><td></td><td></td><td>310</td><td></td><td></td><td></td><td></td><td>315</td><td></td><td></td><td></td><td></td><td>320</td>

<td>Cys</td><td>Arg</td><td>val</td><td>own</td><td>Cheese</td><td>ala</td><td>ala</td><td>phe</td><td>Pro</td><td>ala</td><td>Pro</td><td>How much</td><td>Glu</td><td>lys</td><td>Thr</td><td>How much</td>

<td></td><td></td><td></td><td></td><td>325</td><td></td><td></td><td></td><td></td><td>330</td><td></td><td></td><td></td><td></td><td>335</td><td></td>

<td>Cheese</td><td>lys</td><td>Thr</td><td>Ly3</td><td>Gly</td><td>Arg</td><td>Pro</td><td>lys</td><td>ala</td><td>Pro</td><td>Gin</td><td>val</td><td>Tyr</td><td>Thr</td><td>How much</td><td>Pro</td>

<td></td><td></td><td></td><td>340</td><td></td><td></td><td></td><td></td><td>345</td><td></td><td></td><td></td><td></td><td>350</td><td></td><td></td>

<td>Pro</td><td>Pro</td><td>Ly3</td><td>Glu</td><td>Gin</td><td>Underworld</td><td>ala</td><td>lys</td><td>Asp</td><td>Ly3</td><td>val</td><td>Cheese</td><td>Leu</td><td>Thr</td><td>Cys</td><td>Underworld</td>

<td></td><td></td><td>355</td><td></td><td></td><td></td><td></td><td>360</td><td></td><td></td><td></td><td></td><td>365</td><td></td><td></td><td></td>

<td>How much</td><td>Thr</td><td>A3P</td><td>phe</td><td>phe</td><td>Pro</td><td>Glu</td><td>Asp</td><td>How much</td><td>Thr</td><td>val</td><td>Glu</td><td>Trp</td><td>Gin</td><td>Trp</td><td>own</td>

<td></td><td>370</td><td></td><td></td><td></td><td></td><td>375</td><td></td><td></td><td></td><td></td><td>380</td><td></td><td></td><td></td><td></td>

<td>Gly</td><td>Gin</td><td>Pro</td><td>ala</td><td>Glu</td><td>own</td><td>Tyr</td><td>Lye</td><td>own</td><td>Thr</td><td>Gin</td><td>Pro</td><td>How much</td><td>Underworld</td><td>Asp</td><td>Thr</td>

<td>385</td><td></td><td></td><td></td><td></td><td>390</td><td></td><td></td><td></td><td></td><td>395</td><td></td><td></td><td></td><td></td><td>400</td>

<td>Asp</td><td>Gly</td><td>Cheese</td><td>Tyr</td><td>phe</td><td>How much</td><td>Tyr</td><td>Cheese</td><td>lys</td><td>Leu</td><td>own</td><td>val</td><td>Gin</td><td>lys</td><td>Cheese</td><td>own</td>

<td></td><td></td><td></td><td></td><td>405</td><td></td><td></td><td></td><td></td><td>410</td><td></td><td></td><td></td><td></td><td>415</td><td></td>

<td>Trp</td><td>Glu</td><td>ala</td><td>Gly</td><td>own</td><td>Thr</td><td>phe</td><td>Thr</td><td>Cys</td><td>Cheese</td><td>val</td><td>Leu</td><td>His</td><td>Glu</td><td>Gly</td><td>Leu</td>

<td></td><td></td><td></td><td>420</td><td></td><td></td><td></td><td></td><td>425</td><td></td><td></td><td></td><td></td><td>430</td><td></td><td></td>

<td>His</td><td>own</td><td>His</td><td>His</td><td>Thr</td><td>Glu</td><td>lys</td><td>Cheese</td><td>Leu</td><td>Cheese</td><td>His</td><td>Cheese</td><td>Pro</td><td>Gly</td><td>lys</td><td></td>

<td></td><td></td><td>435</td><td></td><td></td><td></td><td></td><td>440</td><td></td><td></td><td></td><td></td><td>445</td><td></td><td></td><td></td>

<td colspan="3">&lt;210&gt; 141</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

VP / 4572 / AGR

120

EP 2 569 010 B1 <211> 10 <212> PRT <213> Artificial sequence <220>

<223> LCDR1 Ab-11 and Ab-16 <400> 141

Arg Ala Ser Ser Ser Ile Ser Tyr Ile 1 5 <210> 142 <211> 7 <212> PRT <213> Artificial sequence <220>

<223> LCDR2 Ab-11 and Ab-16 <400> 142

Ala Thr Ser Asn Leu Ala Ser 1 5 <210> 143 <211> 9 <212> PRT <213> Artificial sequence <220>

<223> LCDR3 Ab-11 and Ab-16 <400> 143

Gin Gin Trp Cheese Ser Asp Pro Leu Thr 1 5 <210> 144 <211> 5 <212> PRT <213> Artificial sequence <220>

<223> HCDR1 Ab-11 and Ab-16 <400> 144

Asp Tyr Tyr Ile His

5 <210> 145 <211> 17 <212> PRT <213> Artificial sequence <220>

<223> HCDR2 Ab-11 and Ab-16 <400> 145

His

<img file="PL2569010T3_D0005.tif" />

<210> 146 <211> 12 <212> PRT <213> Artificial sequence <220>

<223> HCDR3 Ab-11 and Ab-16 <400> 146

Glu A3p Tyr Asp Gly Thr 1 5 <210> 147 <211> 642 <212> DNA <213> Mus musculus <400> 147 fyr Thr Trp

phe

Pro Tyr

VP / 4572 / AGR

121

EP 2 569 010 B1 caaattgttc tctcccagtc tccagcattc ctgtctgtat ctccagggga taaggtcaca 60 atgacttgca gggccagctc aagtataagt tacatacact ggtttcagca gaagccagga 120 tcctccccca gatcctggat ttatgccaca tccaacctgg cttctggagt ccctggtcgc 180 ttcagtggca gtgggtctgg gacctcttac tctctcacaa tcagcagagt ggaggctgag 240 gatgctgcca cttattactg ccagcagtgg agtagtgacc cactcacgtt cggtgctggg 300 accaagctgg agctgaaacg ggctgatgct gcaccaactg tatccatctt cccaccatcc 360 agtgagcagt taacatctgg aggtgcctca gtcgtgtgct tcttgaacaa cttctacccc 420 aaagacatca atgtcaagtg gaagattgat ggcagtgaac gacaaaatgg cgtcctgaac 480 agttggactg atcaggacag caaagacagc acctacagca tgagcagcac cctcacgttg 540 accaaggacg agtatgaacg acataacagc tatacctgtg aggccactca caagacatca 600 acttcaccca ttgtcaagag cttcaacagg aatgagtgtt ag 642 <210> 148 <211> 213 <212> PRT <213> Mus musculus <400 > 148

VP / 4572 / AGR

122

EP 2 569 010 B1

Gin Ile Val Leu Cheese Gin Cheese Pro Ala Phe Leu Cheese Val Cheese Pro Gly 15 10 15

Asp Lys Val Thr Met Thr Cys Arg Ala Cheese Cheese Ser Ile Cheese Tyr Ile 20 25 30

His Trp Phe Gin Gin Ly3 Pro Gly Cheese Ser Pro Arg Cheese Trp Ile Tyr 35 40 45

Ala Thr Cheese Asn Leu Ala Cheese Gly Val Pro Gly Arg Phe Cheese Gly Cheese 50 55 60

Gly Cheese Gly Thr Cheese Tyr Cheese Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gin Gin Trp Cheese Ser Asp Pro Leu Thr 85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Ala Asp Ala Ala Pro 100 105 110

Thr Val Ser Ile Phe Pro Pro Cheese Ser Glu Gin Leu Thr Cheese Gly Gly 115 120 125

Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp. Asn 130 135 140

Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu Asn 145 150 155 160

Trp cheese Asp Asp. Asp cheese Asp cheese Ly3 Asp Ser Thr Tyr Cheese Met Ser Ser 165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr Thr 180 185 190

Cys Glu Ala Thr His Lye Thr Ser Thr Ser Pro Ile Val Lys Ser Phe 195 200 205

Asn Arg Asn Glu Cys 210 <210> 149 <211> 1338 <212> DNA <213> Mus musculus <400> 149

VP / 4572 / AGR

123

EP 2 569 010 B1

<td>gaagttcagc</td><td>tgcaacagtc</td><td>tggggcagac</td><td>cttgtgcagc</td><td>caggggcctc</td><td>agtcaaggtg</td><td>60</td>

<td>tcctgcacag</td><td>cttctggctt</td><td>cgacattaag</td><td>gactactata</td><td>tacactggat</td><td>gaaacagagg</td><td>120</td>

<td>cctgaccagg</td><td>gcctggagtg</td><td>gattggaagg</td><td>gttgatcctg</td><td>acaatggtga</td><td>gactgaattt</td><td>180</td>

<td>gccccgaagt</td><td>tcccgggcaa</td><td>ggccactttt</td><td>acaacagaca</td><td>catcctccaa</td><td>cacagcctac</td><td>240</td>

<td>ctacaactca</td><td>gaggcctgac</td><td>atctgaggac</td><td>actgccatct</td><td>attactgtgg</td><td>gagagaagac</td><td>300</td>

<td>tacgatggta</td><td>cctacacctg</td><td>gtttccttat</td><td>tggggccaag</td><td>ggactctggt</td><td>cactgtctct</td><td>360</td>

<td>gcagccaaaa</td><td>cgacaccccc</td><td>atctgtctat</td><td>ccactggccc</td><td>ctggatctgc</td><td>tgcccaaact</td><td>420</td>

<td>aactccatgg</td><td>tgaccctggg</td><td>atgcctggtc</td><td>aagggctatt</td><td>tccctgagcc</td><td>agtgacagtg</td><td>480</td>

<td>acctggaact</td><td>ctggatccct</td><td>gtccagcggt</td><td>gtgcacacct</td><td>tcccagctgt</td><td>cctgcagtct</td><td>540</td>

<td>gacctctaca</td><td>ctctgagcag</td><td>ctcagtgact</td><td>gtcccctcca</td><td>gcacctggcc</td><td>cagcgagacc</td><td>600</td>

<td>gtcacctgca</td><td>acgttgccca</td><td>cccggccagc</td><td>agcaccaagg</td><td>tggacaagaa</td><td>aattgtgccc</td><td>660</td>

<td>agggattgtg</td><td>gttgtaagcc</td><td>ttgcatatgt</td><td>acagtcccag</td><td>aagtatcatc</td><td>tgtcttcatc</td><td>720</td>

<td>ttccccccaa</td><td>agcccaagga</td><td>tgtgctcacc</td><td>attactctga</td><td>ctcctaaggt</td><td>cacgtgtgtt</td><td>780</td>

<td>gtggtagaca</td><td>tcagcaagga</td><td>tgatcccgag</td><td>gtccagttca</td><td>gctggtttgt</td><td>agatgatgtg</td><td>840</td>

<td>gaggtgcaca</td><td>cagctcagac</td><td>gcaaccccgg</td><td>gaggagcagt</td><td>tcaacagcac</td><td>tttccgctca</td><td>900</td>

<td>gtcagtgaac</td><td>ttcccatcat</td><td>gcaccaggac</td><td>tggctcaatg</td><td>gcaaggagtt</td><td>caaatgcagg</td><td>960</td>

<td>gtcaacagtg</td><td>cagctttccc</td><td>tgcccccatc</td><td>gagaaaacca</td><td>tctccaaaac</td><td>caaaggcaga</td><td>1020</td>

<td>ccgaaggctc</td><td>cacaggtgta</td><td>caccattcca</td><td>cctcccaagg</td><td>agcagatggc</td><td>caaggataaa</td><td>1080</td>

<td>gtcagtctga</td><td>cctgcatgat</td><td>aacagacttc</td><td>ttccctgaag</td><td>acattactgt</td><td>ggagtggcag</td><td>1140</td>

<td>tggaatgggc</td><td>agccagcgga</td><td>gaactacaag</td><td>aacactcagc</td><td>ccatcatgga</td><td>cacagatggc</td><td>1200</td>

<td>tcttacttca</td><td>tctacagcaa</td><td>gctcaatgtg</td><td>cagaagagca</td><td>actgggaggc</td><td>aggaaatact</td><td>1260</td>

<td>ttcacctgct</td><td>ctgtgttaca</td><td>tgagggcctg</td><td>cacaaccacc</td><td>atactgagaa</td><td>gagcctctcc</td><td>1320</td>

<td>cactctcctg</td><td>gtaaatga</td><td></td><td></td><td></td><td></td><td>1338</td>

<210> 150 <211> 445 <212> PRT <213> Mus musculus <400> 150

VP / 4572 / AGR

124

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Cheese Gly Ala Asp Leu Val Gin Pro Gly Ala 15 10 15

Ser Val Lys Val Ser Cys Thr Ala Ser Gly Phe Asp Ile Lys Asp Tyr 20 25 30

Tyr Ile Trp Met Lys Gin Arg Pro Asp Gin Gly Leu Glu Trp Ile 35 40 45

Gly Arg Asp Asp Pro Asp Asn Gly Glu Thr Glu Phe Ala Pro Lys Phe 50 55 60

Pro Gly Lys Ala Thr Phe Thr Thr Asp Thr Cheese Ser Asn Thr Ala Tyr 65 70 75 80

Leu Gin Leu Arg Gly Leu Thr Ser Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95

Gly Arg Glu Asp Tyr Asp Gly Thr Tyr Thr Trp Phe Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser 115 115 125

Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr Asn Cheese Met Val 130 135 140

Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Thr Trp Asn Cheese Gly Cheese Leu Cheese Cheese Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Cheese Asp Leu Tyr Thr Leu Cheese Cheese Cheese Val Thr Val Pro 180 185 190

Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro 195 200 205

Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp. Cys Gly 210 215 220

VP / 4572 / AGR

125

EP 2 569 010 B1

Cys Ly3 Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile 225 230 235 240

Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys 245 250 255

Val Thr Cys Val Val Val Asp Ile Ile Lys Asp A3p Pro Glu Val Gin 260 265 270

Phe Ser Trp Phe Val Asp Asp. Val. Glu. Val. His Thr Ala Gin Thr. Gin 275 280 285

Pro Arg Glu Glu Gin Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu 290 295 300

Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg 305 310 315 320

Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Ile Glu Ly3 Thr Ile Ser Lys 325 330 335

Thr Lys Gly Arg Pro Lys Ala Pro Gin Val Tyr Thr How much Pro Pro Pro 340 345 350

Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr 355 360 365

Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn Gly Gin 370 375 380

Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro How Much Met Asp Thr Asp Gly 385 390 395 400

Cheese Tyr Phe Ile Tyr Cheese Lys Leu Asn Val Gin Lys Cheese Asn Trp Glu 405 410 415

Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn 420 425 430

His His Thr Glu Lys Ser Leu Ser His Cheese Pro Gly Lys 435 440 445 <210> 151 <211> 318 <212> DNA <213> Mus musculus <400> 151

VP / 4572 / AGR

126

EP 2 569 010 B1

<td>gacatccagt</td><td>tgacccagtc</td><td>tccatccttc</td><td>ctgtctgcat</td><td>ctgtaggaga</td><td>cagagtcacc</td><td>60</td>

<td>atcacttgca</td><td>gggccagctc</td><td>aagtataagt</td><td>tacatacact</td><td>ggtatcagca</td><td>aaaaccaggg</td><td>120</td>

<td>aaagccccta</td><td>agctcctgat</td><td>ctatgccaca</td><td>tccaacctgg</td><td>cttctggggt</td><td>cccatcaagg</td><td>180</td>

<td>ttcagcggca</td><td>gtggatctgg</td><td>gacagaattc</td><td>actctcacaa</td><td>tcagcagcct</td><td>gcagcctgaa</td><td>240</td>

<td>gattttgcaa</td><td>cttattactg</td><td>tcagcagtgg</td><td>agtagtgacc</td><td>cactcacgtt</td><td>cggcggaggg</td><td>300</td>

<td>accaaggtgg</td><td>agatcaaa</td><td></td><td></td><td></td><td></td><td>318</td>

<210> 152 <211> 106 <212> PRT <213> Mus musculus

<td colspan="4">&lt;400&gt; 152</td><td rowspan="2">Thr 5</td><td rowspan="2">Gin</td><td rowspan="2">Cheese</td><td rowspan="2">Pro</td><td rowspan="2">Cheese</td><td rowspan="2">phe 10</td><td rowspan="2">Leu</td><td rowspan="2">Cheese</td><td rowspan="2">ala</td><td rowspan="2">Cheese</td><td rowspan="2">val 15</td><td rowspan="2">Gly</td>

<td>Asp 1</td><td>How much</td><td>Gin</td><td>Leu</td>

<td>A3P</td><td>Arg</td><td>val</td><td>Thr</td><td>How much</td><td>Thr</td><td>Cys</td><td>Arg</td><td>ala</td><td>Cheese</td><td>Cheese</td><td>Cheese</td><td>How much</td><td>Cheese</td><td>Tyr</td><td>How much</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>thirty</td><td></td><td></td>

<td>His</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>lys</td><td>Pro</td><td>Gly</td><td>lys</td><td>ala</td><td>Pro</td><td>lys</td><td>Leu</td><td>Leu</td><td>How much</td><td>Tyr</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>ala</td><td>Thr</td><td>Cheese</td><td>own</td><td>Leu</td><td>ala</td><td>Cheese</td><td>Gly</td><td>val</td><td>Pro</td><td>Cheese</td><td>Arg</td><td>phe</td><td>Cheese</td><td>Gly</td><td>Cheese</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Gly</td><td>Cheese</td><td>Gly</td><td>Thr</td><td>Glu</td><td>phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>How much</td><td>Cheese</td><td>Cheese</td><td>Leu</td><td>Gin</td><td>Pro</td><td>Glu</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Asp</td><td>phe</td><td>ala</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Trp</td><td>Cheese</td><td>Cheese</td><td>Asp</td><td>Pro</td><td>Leu</td><td>Thr</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>lys</td><td>val</td><td>Glu</td><td>How much</td><td>Ly3</td><td></td><td></td><td></td><td></td><td></td><td></td>

100 105 <210> 153 <211> 363 <212> DNA <213> Mus musculus <400> 153

<td>gaggtgcagc</td><td>tggtgcagtc</td><td>tggggctgag</td><td>gtgaagaagc</td><td>ctggggcctc</td><td>agtgaaggtc</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggatt</td><td>cgacattaag</td><td>gactactata</td><td>tacactgggt</td><td>gcgacaggcc</td><td>120</td>

<td>cctggacaag</td><td>ggcttgagtg</td><td>gatcggaagg</td><td>gttgatcctg</td><td>acaatggtga</td><td>gactgaattt</td><td>180</td>

<td>gccccgaagt</td><td>tcccgggcaa</td><td>ggtcaccatg</td><td>accacagaca</td><td>cgtccatcag</td><td>cacagcctac</td><td>240</td>

<td>atggagctga</td><td>gcaggctgag</td><td>atctgacgac</td><td>acggccgtgt</td><td>attactgtgc</td><td>gagagaagac</td><td>300</td>

<td>tacgatggta</td><td>cctacacctg</td><td>gtttccttat</td><td>tggggccaag</td><td>ggactctggt</td><td>caccgtctct</td><td>360</td>

<td>agt</td><td></td><td></td><td></td><td></td><td></td><td>363</td>

<210> 154 <211> 121 <212> PRT <213> Mus musculus <400> 154

VP / 4572 / AGR

127

EP 2 569 010 B1

Glu Val Gin Leu Val Gin Cheese Gly Ala Glu Val Lys Pro Gly Ala 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Asp Ile Lys Asp Tyr 20 25 30

Tyr Ile His Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Ile 35 40 45

Gly Arg Asp Asp Pro Asp Asn Gly Glu Thr Glu Phe Ala Pro Lys Phe 50 55 60

Pro Gly Lys Val Thr Met Thr Thr Asp Thr Ser Ile Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Arg Leu Arg Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Glu Asp Tyr Asp Gly Thr Tyr Thr Trp Phe Pro Tyr Trp Gly 100 105 110

Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 155 <211> 639 <212> DNA <213> Artificial sequence <220>

<223> Sequence of humanized antibody <400> 155

<td>gacatccagt</td><td>tgacccagtc</td><td>tccatccttc</td><td>ctgtctgcat</td><td>ctgtaggaga</td><td>cagagtcacc</td><td>60</td>

<td>atcacttgca</td><td>gggccagctc</td><td>aagtataagt</td><td>tacatacact</td><td>ggtatcagca</td><td>aaaaccaggg</td><td>120</td>

<td>aaagccccta</td><td>agctcctgat</td><td>ctatgccaca</td><td>tccaacctgg</td><td>cttctggggt</td><td>cccatcaagg</td><td>180</td>

<td>ttcagcggca</td><td>gtggatctgg</td><td>gacagaattc</td><td>actctcacaa</td><td>tcagcagcct</td><td>gcagcctgaa</td><td>240</td>

<td>gattttgcaa</td><td>cttattactg</td><td>tcagcagtgg</td><td>agtagtgacc</td><td>cactcacgtt</td><td>cggcggaggg</td><td>300</td>

<td>accaaggtgg</td><td>agatcaaacg</td><td>tacggtggct</td><td>gcaccatctg</td><td>tcttcatctt</td><td>cccgccatct</td><td>360</td>

<td>gatgagcagt</td><td>tgaaatctgg</td><td>aactgcctct</td><td>gttgtgtgcc</td><td>tgctgaataa</td><td>cttctatccc</td><td>420</td>

<td>agagaggcca</td><td>aagtacagtg</td><td>gaaggtggat</td><td>aacgccctcc</td><td>aatcgggtaa</td><td>ctcccaggag</td><td>480</td>

<td>agtgtcacag</td><td>agcaggacag</td><td>caaggacagc</td><td>acctacagcc</td><td>tcagcagcac</td><td>cctgacgctg</td><td>540</td>

<td>agcaaagcag</td><td>actacgagaa</td><td>acacaaagtc</td><td>tacgcctgcg</td><td>aagtcaccca</td><td>tcagggcctg</td><td>600</td>

<td>agctcgcccg</td><td>tcacaaagag</td><td>cttcaacagg</td><td>ggagagtgt</td><td></td><td></td><td>639</td>

<210> 156 <211> 213 <212> PRT <213> Artificial sequence <220>

<223> Sequence of humanized antibody <400> 156

VP / 4572 / AGR

128

EP 2 569 010 B1

Asp Ile Gin Leu Thr 1 5

Gin Cheese Pro Cheese Phe 10

Asp Arg Val Thr Ile 20

Thr Cys Arg Ala Ser 25

His Trp Tyr Gin Gin 35

Lys Pro Gly Lys Ala 40

Ala Thr Cheese Asn Leu 50

Ala Ser Gly Val Pro 55

Gly Cheese Gly Thr Glu 65

Phe Thr Leu Thr Ile 70

Asp Phe Ala Thr Tyr 85

Tyr Cys Gin Gin Trp 90

Phe Gly Gly Gly Thr 100

Lys Val Glu Ile Lys 105

Ser Val Phe Ile Phe 115

Pro Pro Ser Asp Glu 120

Ala Ser Val Val Cys 130

Leu Leu Asn Asn Phe 135

Val Gin Trp Lys Val 145

Asp Asn Ala Leu Gin 150

Ser Val Thr Glu Gin 165

Asp Ser Lys Asp Ser 170

Thr Leu Thr Leu Ser 180

Lys Ala Asp Tyr Glu 185

Cys Glu Val Thr His 195

Gin Gly Leu Cheese Ser 200

Leu

Cheese

Pro

Cheese

Cheese

Cheese

Arg

Gin

Tyr

Cheese

155

Thr

Lye

Pro

Cheese Ala Ser Val Gly 15

Ser Ile Ser Tyr Ile 30

Lys Leu Leu Ile Tyr 45

Arg Phe Cheese Gly Ser 60

Leu Gin Pro Glu 80 cheese

Asp Pro Leu Thr 95 cheese

Thr Val Ala Ala Pro 110

Leu Lys Ser Gly Thr 125

Pro Arg Glu Ala Lys 140

Gly Asn Cheese Gin Glu 160

Tyr Ser Leu Cheese Ser 175

His Lys Val Tyr Ala 190

Val Thr Lys Ser Phe 205

Asn Arg Gly Glu Cys 210 <210> 157 <211> 1341 <212> DNA <213> Artificial sequence <220>

<223> Sequence of humanized antibody <400> 157

VP / 4572 / AGR

129

EP 2 569 010 B1

<td>gaggtgcagc</td><td>tggtgcagtc</td><td>tggggctgag</td><td>gtgaagaagc</td><td>ctggggcctc</td><td>agtgaaggtc</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggatt</td><td>cgacattaag</td><td>gactactata</td><td>tacactgggt</td><td>gcgacaggcc</td><td>120</td>

<td>cctggacaag</td><td>ggcttgagtg</td><td>gatcggaagg</td><td>gttgatcctg</td><td>acaatggtga</td><td>gactgaattt</td><td>180</td>

<td>gccccgaagt</td><td>tcccgggcaa</td><td>ggtcaccatg</td><td>accacagaca</td><td>cgtccatcag</td><td>cacagcctac</td><td>240</td>

<td>atggagctga</td><td>gcaggctgag</td><td>atctgacgac</td><td>acggccgtgt</td><td>attactgtgc</td><td>gagagaagac</td><td>300</td>

<td>tacgatggta</td><td>cctacacctg</td><td>gtttccttat</td><td>tggggccaag</td><td>ggactctggt</td><td>caccgtctct</td><td>360</td>

<td>agtgcctcca</td><td>ccaagggccc</td><td>atcggtcttc</td><td>cccctggcgc</td><td>cctgctccag</td><td>gagcacctcc</td><td>420</td>

<td>gagagcacag</td><td>cggccctggg</td><td>ctgcctggtc</td><td>aaggactact</td><td>tccccgaacc</td><td>ggtgacggtg</td><td>480</td>

<td>tcgtggaact</td><td>caggcgctct</td><td>gaccagcggc</td><td>gtgcacacct</td><td>tcccagctgt</td><td>cctacagtcc</td><td>540</td>

<td>tcaggactct</td><td>actccctcag</td><td>cagcgtggtg</td><td>accgtgccct</td><td>ccagcaactt</td><td>cggcacccag</td><td>600</td>

<td>acctacacct</td><td>gcaacgtaga</td><td>tcacaagccc</td><td>agcaacacca</td><td>aggtggacaa</td><td>gacagttgag</td><td>660</td>

<td>cgcaaatgtt</td><td>gtgtcgagtg</td><td>cccaccgtgc</td><td>ccagcaccac</td><td>ctgtggcagg</td><td>accgtcagtc</td><td>720</td>

<td>ttcctcttcc</td><td>ccccaaaacc</td><td>caaggacacc</td><td>ctcatgatct</td><td>cccggacccc</td><td>tgaggtcacg</td><td>780</td>

<td>tgcgtggtgg</td><td>tggacgtgag</td><td>ccacgaagac</td><td>cccgaggtcc</td><td>agttcaactg</td><td>gtacgtggac</td><td>840</td>

<td>ggcgtggagg</td><td>tgcataatgc</td><td>caagacaaag</td><td>ccacgggagg</td><td>agcagttcaa</td><td>cagcacgttc</td><td>900</td>

<td>cgtgtggtca</td><td>gcgtcctcac</td><td>cgttgtgcac</td><td>caggactggc</td><td>tgaacggcaa</td><td>ggagtacaag</td><td>960</td>

<td>tgcaaggtct</td><td>ccaacaaagg</td><td>cctcccagcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>gggcagcccc</td><td>gagaaccaca</td><td>ggtgtacacc</td><td>ctgcccccat</td><td>cccgggagga</td><td>gatgaccaag</td><td>1080</td>

<td>aaccaggtca</td><td>gcctgacctg</td><td>cctggtcaaa</td><td>ggcttctacc</td><td>ccagcgacat</td><td>cgccgtggag</td><td>1140</td>

<td>tgggagagca</td><td>atgggcagcc</td><td>ggagaacaac</td><td>tacaagacca</td><td>cacctcccat</td><td>gctggactcc</td><td>1200</td>

<td>gacggctcct</td><td>tcttcctcta</td><td>cagcaagctc</td><td>accgtggaca</td><td>agagcaggtg</td><td>gcagcagggg</td><td>1260</td>

<td>aacgtcttct</td><td>catgctccgt</td><td>gatgcatgag</td><td>gctctgcaca</td><td>accactacac</td><td>gcagaagagc</td><td>1320</td>

<td>ctctccctgt</td><td>ctccgggtaa</td><td>and</td><td></td><td></td><td></td><td>1341</td>

<210> 158 <211> 447 <212> PRT <213> Artificial sequence <220>

<223> Sequence of humanized antibody <400> 158

EP 2 569 010 B1

<td colspan="2">VP / 4572 / AGR</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td colspan="2">130</td><td></td><td></td><td></td><td></td><td></td>

<td>Glu</td><td>Val</td><td>Gin</td><td>Leu</td><td>Val</td><td>Gin</td><td>Ser</td><td>Gly</td><td>Ala</td><td>Glu</td><td>Val</td><td>Lys</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Ala</td>

<td>1</td><td></td><td></td><td></td><td>5</td><td></td><td></td><td></td><td></td><td>10</td><td></td><td></td><td></td><td></td><td>15</td><td></td>

<td>Ser</td><td>Val</td><td>Lys</td><td>Val</td><td>Ser</td><td>Cys</td><td>Lys</td><td>Ala</td><td>Ser</td><td>Gly</td><td>Phe</td><td>Asp</td><td>Ile</td><td>Lys</td><td>Asp</td><td>Tyr</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>30</td><td></td><td></td>

<td>Tyr</td><td>ile</td><td>His</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Gly</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Ile</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Gly</td><td>Arg</td><td>Val</td><td>Asp</td><td>Pro</td><td>Asp</td><td>Asn</td><td>Gly</td><td>Glu</td><td>Thr</td><td>Glu</td><td>Phe</td><td>Ala</td><td>Pro</td><td>Lys</td><td>Phe</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Pro</td><td>Gly</td><td>Lys</td><td>Val</td><td>Thr</td><td>Met</td><td>Thr</td><td>Thr</td><td>Asp</td><td>Thr</td><td>Ser</td><td>Ile</td><td>Ser</td><td>Thr</td><td>Ala</td><td>Tyr</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Met</td><td>Glu</td><td>Leu</td><td>Ser</td><td>Arg</td><td>Leu</td><td>Arg</td><td>Ser</td><td>Asp</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>Ala</td><td>Arg</td><td>Glu</td><td>Asp</td><td>Tyr</td><td>Asp</td><td>Gly</td><td>Thr</td><td>Tyr</td><td>Thr</td><td>Trp</td><td>Phe</td><td>Pro</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Leu</td><td>Val</td><td>Thr</td><td>Val</td><td>Ser</td><td>Ser</td><td>Ala</td><td>Ser</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Ser</td>

PZ/4572/AGR

131

EP 2 569 010 B1

115 120 125

Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 Π0 175

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Asn Phe Gly Thr Gin Thr Tyr Thr Cy3 Asn Val Asp His 195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys 210 215 220

Val Glu Cys Pro Pro Cy3 Pro Ala Pro Pro Val Ala Gly Pro Ser Val 225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270

Val Gin Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Ly3 275 280 285

Thr Lys Pro Arg Glu Glu Gin Phe Asn Ser Thr Phe Arg Val Val Ser 290 295 300

Val Leu Thr Val Val His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Ly3 Thr Ile 325 330 335

Ser Lys Thr Lys Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro 340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gin Val Ser Leu Thr Cys Leu

PZ/4572/AGR

132

EP 2 569 010 B1

355

360

365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380

Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser 385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415

Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430

His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 159 <211> 11 <212> PRT <213> Sekwencja sztuczna <220>

<223> LCDR1 Ab-12 <400> 159

Arg Ala Ser Gin Asp Ile Ser Asn Tyr Leu Asn

10 <210> 160 <211> 7 <212> PRT <213> Sekwencja sztuczna <220>

<223> LCDR2 Ab-12 <400> 160

Tyr Thr Ser Thr Leu Gin Ser

5 <210> 161 <211> 9 <212> PRT <213> Sekwencja sztuczna <220>

<223> LCDR3 Ab-12 <400> 161

Gin Gin Gly Asp Thr Leu Pro Tyr Thr

5 <210> 162 <211> 5 <212> PRT <213> Sekwencja sztuczna <220>

<223> HCDR1 Ab-12 <400> 162

Asp Tyr Asn Met His 1 5 <210> 163 <211> 17 <212> PRT <213> Sekwencja sztuczna <220>

<223> HCDR2 Ab-12 <400> 163

PZ/4572/AGR

133

EP 2 569 010 B1

Glu Ile Asn Pro Asn Ser Gly Gly Ser Gly Tyr Asn Gin Lys Phe Ly 15 10 15

Gly <210> 164 <211> 14 <212> PRT <213> Sekwencja sztuczna <220>

<223> HCDR3 Ab-12 <400> 164

Leu Gly Tyr Tyr Gly Aen Tyr Glu Asp Trp Tyr Phe A3p Val 15 10 <210> 165 <211> 645 <212> DNA <213> Mus musculus <400> 165

<td>gatctccaga</td><td>tgacacagac</td><td>tacttcctcc</td><td>ctgtctgcct</td><td>ctctgggaga</td><td>cagagtcacc</td><td>60</td>

<td>atcagttgca</td><td>gggcaagtca</td><td>ggacattagc</td><td>aattatttaa</td><td>actggtatca</td><td>gcagaaacca</td><td>120</td>

<td>gatggaactg</td><td>ttaagctcct</td><td>gatcttctac</td><td>acatcaacat</td><td>tacagtcagg</td><td>agtcccatcg</td><td>180</td>

<td>aggttcagtg</td><td>gcagtgggtc</td><td>tggaacaaat</td><td>tattctctca</td><td>ccattaccaa</td><td>cctggagcaa</td><td>240</td>

<td>gatgatgctg</td><td>ccacttactt</td><td>ttgccaacag</td><td>ggtgatacgc</td><td>ttccgtacac</td><td>gttcggaggg</td><td>300</td>

<td>gggaccaagc</td><td>tggaaataaa</td><td>acgggctgat</td><td>gctgcaccaa</td><td>ctgtatccat</td><td>cttcccacca</td><td>360</td>

<td>tccagtgagc</td><td>agttaacatc</td><td>tggaggtgcc</td><td>tcagtcgtgt</td><td>gcttcttgaa</td><td>caacttctac</td><td>420</td>

<td>cccaaagaca</td><td>tcaatgtcaa</td><td>gtggaagatt</td><td>gatggcagtg</td><td>aacgacaaaa</td><td>tggcgtcctg</td><td>480</td>

<td>aacagttgga</td><td>ctgatcagga</td><td>cagcaaagac</td><td>agcacctaca</td><td>gcatgagcag</td><td>caccctcacg</td><td>540</td>

<td>ttgaccaagg</td><td>acgagtatga</td><td>acgacataac</td><td>agctatacct</td><td>gtgaggccac</td><td>tcacaagaca</td><td>600</td>

<td>tcaacttcac</td><td>ccattgtcaa</td><td>gagcttcaac</td><td>aggaatgagt</td><td>gttag</td><td></td><td>645</td>

<210> 166 <211> 214 <212> PRT <213> Mus musculus <400> 166

PZ/4572/AGR

134

EP 2 569 010 B1

Asp Leu Gin Met Thr Gin Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 15 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gin Asp Ile Ser Asn Tyr 20 25 30

Leu Asn Trp Tyr Gin Gin Lys Pro Asp Gly Thr Val Lys Leu Leu Ile 35 40 45

Phe Tyr Thr Ser Thr Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asn Tyr Ser Leu Thr Ile Thr A3n Leu Glu Gin 65 70 75 80

Asp Asp Ala Ala Thr Tyr Phe Cys Gin Gin Gly Asp Thr Leu Pro Tyr 85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110

Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gin Leu Thr Ser Gly 115 120 125

Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140

Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gin Asn Gly Val Leu 145 150 155 160

Asn Ser Trp Thr A3p Gin A3p Ser Lys Aep Ser Thr Tyr Ser Met Ser 165 170 175

Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr 180 185 190

Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205

Phe Asn Arg Asn Glu Cys 210 <210> 167 <211> 1344 <212> DNA <213> Mus musculus <400> 167

PZ/4572/AGR

135

EP 2 569 010 B1

<td>gaggtccagt</td><td>tgcaacagtc</td><td>tggacctgaa</td><td>ctaatgaagc</td><td>ctggggcttc</td><td>agtgaagatg</td><td>60</td>

<td>tcctgcaagg</td><td>cttctggata</td><td>cacattcact</td><td>gactacaaca</td><td>tgcactggat</td><td>gaagcagaac</td><td>120</td>

<td>caaggaaaga</td><td>gcctagagtg</td><td>gataggagag</td><td>attaatccta</td><td>acagtggtgg</td><td>ttctggttac</td><td>180</td>

<td>aaccagaagt</td><td>tcaaaggcaa</td><td>ggccacattg</td><td>actgtagaca</td><td>agtcctccag</td><td>cacagcctac</td><td>240</td>

<td>atggagctcc</td><td>gcagcctgac</td><td>atctgaggac</td><td>tctgcagtct</td><td>attactgtgc</td><td>aagattgggc</td><td>300</td>

<td>tactatggta</td><td>actacgagga</td><td>ctggtatttc</td><td>gatgtctggg</td><td>gcgcagggac</td><td>cacggtcacc</td><td>360</td>

<td>gtctcctctg</td><td>ccaaaacgac</td><td>acccccatct</td><td>gtctatccac</td><td>tggcccctgg</td><td>atctgctgcc</td><td>420</td>

<td>caaactaact</td><td>ccatggtgac</td><td>cctgggatgc</td><td>ctggtcaagg</td><td>gctatttccc</td><td>tgagccagtg</td><td>480</td>

<td>acagtgacct</td><td>ggaactctgg</td><td>atccctgtcc</td><td>agcggtgtgc</td><td>acaccttccc</td><td>agctgtcctg</td><td>540</td>

<td>cagtctgacc</td><td>tctacactct</td><td>gagcagctca</td><td>gtgactgtcc</td><td>cctccagcac</td><td>ctggcccagc</td><td>600</td>

<td>gagaccgtca</td><td>cctgcaacgt</td><td>tgcccacccg</td><td>gccagcagca</td><td>ccaaggtgga</td><td>caagaaaatt</td><td>660</td>

<td>gtgcccaggg</td><td>attgtggttg</td><td>taagccttgc</td><td>atatgtacag</td><td>tcccagaagt</td><td>atcatctgtc</td><td>720</td>

<td>ttcatcttcc</td><td>ccccaaagcc</td><td>caaggatgtg</td><td>ctcaccatta</td><td>ctctgactcc</td><td>taaggtcacg</td><td>780</td>

<td>tgtgttgtgg</td><td>tagacatcag</td><td>caaggatgat</td><td>cccgaggtcc</td><td>agttcagctg</td><td>gtttgtagat</td><td>840</td>

<td>gatgtggagg</td><td>tgcacacagc</td><td>tcagacgcaa</td><td>ccccgggagg</td><td>agcagttcaa</td><td>cagcactttc</td><td>900</td>

<td>cgctcagtca</td><td>gtgaacttcc</td><td>catcatgcac</td><td>caggactggc</td><td>tcaatggcaa</td><td>ggagttcaaa</td><td>960</td>

<td>tgcagggtca</td><td>acagtgcagc</td><td>tttccctgcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>ggcagaccga</td><td>aggctccaca</td><td>ggtgtacacc</td><td>attccacctc</td><td>ccaaggagca</td><td>gatggccaag</td><td>1080</td>

<td>gataaagtca</td><td>gtctgacctg</td><td>catgataaca</td><td>gacttcttcc</td><td>ctgaagacat</td><td>tactgtggag</td><td>1140</td>

<td>tggcagtgga</td><td>atgggcagcc</td><td>agcggagaac</td><td>tacaagaaca</td><td>ctcagcccat</td><td>catggacaca</td><td>1200</td>

<td>gatggctctt</td><td>acttcatcta</td><td>cagcaagctc</td><td>aatgtgcaga</td><td>agagcaactg</td><td>ggaggcagga</td><td>1260</td>

<td>aatactttca</td><td>cctgctctgt</td><td>gttacatgag</td><td>ggcctgcaca</td><td>accaccatac</td><td>tgagaagagc</td><td>1320</td>

ctctcccact ctcctggtaa atga 1344 <210> 168 <211> 447 <212> PRT <213> Mus musculus <400> 168

PZ/4572/AGR

136

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Met Lys Pro Gly Ala 15 10 15

Ser Val Lye Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr A3p Tyr 20 25 30

Asn Met His Trp Met Lys Gin Asn Gin Gly Lys Ser Leu Glu Trp Ile 35 40 45

Gly Glu Ile Asn Pro Asn Ser Gly Gly Ser Gly Tyr Asn Gin Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Gly Tyr Tyr Gly Asn Tyr Glu Asp Trp Tyr Phe Asp Val 100 105 110

Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Ala Lys Thr Thr Pro 115 120 125

Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr Asn Ser 130 135 140

Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val

PZ/4572/AGR

137

EP 2 569 010 B1

145 150 155 160

Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe 165 170 175

Pro Ala Val Leu Gin Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr 180 185 190

Val Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala 195 200 205

His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp 210 215 220

Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val 225 230 235 240

Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr 245 250 255

Pro Lys Val Thr Cys Val Val Val Asp Ile Ser Lye Asp Asp Pro Glu 260 265 270

Val Gin Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gin 275 280 285

Thr Gin Pro Arg Glu Glu Gin Phe Asn Ser Thr Phe Arg Ser Val Ser 290 295 300

Glu Leu Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Lys 305 310 315 320

Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335

Ser Lys Thr Lys Gly Arg Pro Lys Ala Pro Gin Val Tyr Thr Ile Pro 340 345 350

Pro Pro Lys Glu Gin Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met 355 360 365

Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn 370 375 380

Gly Gin Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro Ile Met Asp Thr

PZ/4572/AGR

138

EP 2 569 010 B1

385 390 395 400

Asp Gly Ser Tyr Phe Ile Tyr Ser Lys Leu Asn Val Gin Lys Ser Asn 405 410 415

Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu 420 425 430

Hi3 Asn Hi3 His Thr Glu Lys Ser Leu Ser Hi3 Ser Pro Gly Lys 435 440 445 <210> 169 <211> 12 <212> PRT <213> Sekwencja sztuczna <220>

<223> LCDR1 Ab-13 i Ab-14 <400> 169

Arg Ala Ser Ser Ser Val Thr Ser Ser Tyr Leu Asn 15 10 <210> 170 <211> 7 <212> PRT <213> Sekwencja sztuczna <220>

<223> LCDR2 Ab-13 i Ab-14 <400> 170

Ser Thr Ser Asn Leu Ala Ser

5 <210> 171 <211> 9 <212> PRT <213> Sekwencja sztuczna <220>

<223> LCDR3 Ab-13 i Ab-14 <400> 171

Gin Gin Tyr Asp Phe Phe Pro Ser Thr 1 5 <210> 172 <211> 5 <212> PRT <213> Sekwencja sztuczna <220>

<223> HCDR1 Ab-13 i Ab-14 <400> 172

Asp Tyr Tyr Met Asn

5 <210> 173 <211> 17 <212> PRT <213> Sekwencja sztuczna <220>

<223> HCDR2 Ab-13 i Ab-14 <400> 173

Asp Ile Asn Pro Tyr Asn A3p Asp Thr Thr Tyr Asn His Lys Phe Lys 15 10 15

Gly <210> 174 <211> 11 <212> PRT <213> Sekwencja sztuczna

PZ/4572/AGR

139

EP 2 569 010 B1 <220>

<223> HCDR3 Ab-13 i Ab-14 <400> 174

Glu Thr Ala Val Ile Thr Thr Asn Ala Met Asp 15 10 <210> 175 <211> 645 <212> DNA <213> Mus musculus <400> 175

<td>cagattgttc</td><td>tcacccagtc</td><td>tccagcaatc</td><td>atgtctgcat</td><td>ctccagggga</td><td>gaaggtcacc</td><td>60</td>

<td>atgacctgca</td><td>gggccagctc</td><td>aagtgtaact</td><td>tccagttact</td><td>tgaactggta</td><td>ccagcagaag</td><td>120</td>

<td>ccaggatctt</td><td>cccccaaact</td><td>ctggatttat</td><td>agcacatcca</td><td>acctggcttc</td><td>aggagtccca</td><td>180</td>

<td>gctcgcttca</td><td>gtggcagtgg</td><td>gtctgggacc</td><td>tcttactctc</td><td>tcacaatcag</td><td>cagtgtggag</td><td>240</td>

<td>gctgaggatg</td><td>ctgccactta</td><td>ttactgccag</td><td>cagtatgatt</td><td>ttttcccatc</td><td>gacgttcggt</td><td>300</td>

<td>ggaggcacca</td><td>agctggaaat</td><td>caagcgggct</td><td>gatgctgcac</td><td>caactgtatc</td><td>catcttccca</td><td>360</td>

<td>ccatccagtg</td><td>agcagttaac</td><td>atctggaggt</td><td>gcctcagtcg</td><td>tgtgcttctt</td><td>gaacaacttc</td><td>420</td>

<td>taccccaaag</td><td>acatcaatgt</td><td>caagtggaag</td><td>attgatggca</td><td>gtgaacgaca</td><td>aaatggcgtc</td><td>480</td>

<td>ctgaacagtt</td><td>ggactgatca</td><td>ggacagcaaa</td><td>gacagcacct</td><td>acagcatgag</td><td>cagcaccctc</td><td>540</td>

<td>acgttgacca</td><td>aggacgagta</td><td>tgaacgacat</td><td>aacagctata</td><td>cctgtgaggc</td><td>cactcacaag</td><td>600</td>

<td>acatcaactt</td><td>cacccatcgt</td><td>caagagcttc</td><td>aacaggaatg</td><td>agtgt</td><td></td><td>645</td>

<210> 176 <211> 215 <212> PRT <213> Mus musculus <400> 176

PZ/4572/AGR 140

EP 2 569 010 B1

<td>Gin 1</td><td>Ile Val</td><td>Leu</td><td>Thr 5</td><td>Gin</td><td>Ser</td><td>Pro</td><td>Ala</td><td>Ile Met 10</td><td>Ser</td><td>Ala</td><td>Ser</td><td>Pro 15</td><td>Gly</td>

<td>Glu</td><td>Lys Val</td><td>Thr</td><td>Met</td><td>Thr</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Ser Ser</td><td>Ser</td><td>Val</td><td>Thr</td><td>Ser</td><td>Ser</td>

<td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td>30</td><td></td><td></td>

<td>Tyr</td><td>Leu Asn</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly Ser</td><td>Ser</td><td>Pro</td><td>Lys</td><td>Leu</td><td>Trp</td>

<td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Ile</td><td>Tyr Ser</td><td>Thr</td><td>Ser</td><td>Asn</td><td>Leu</td><td>Ala</td><td>Ser</td><td>Gly Val</td><td>Pro</td><td>Ala</td><td>Arg</td><td>Phe</td><td>Ser</td>

<td></td><td>50</td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Gly</td><td>Ser Gly</td><td>Ser</td><td>Gly</td><td>Thr</td><td>Ser</td><td>Tyr</td><td>Ser</td><td>Leu Thr</td><td>Ile</td><td>Ser</td><td>Ser</td><td>Val</td><td>Glu</td>

<td>65</td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Ala</td><td>Glu Asp</td><td>Ala</td><td>Ala</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin Gin</td><td>Tyr</td><td>Asp</td><td>Phe</td><td>Phe</td><td>Pro</td>

<td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td>95</td><td></td>

<td>Ser</td><td>Thr Phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>Lys</td><td>Leu</td><td>Glu Ile</td><td>Lys</td><td>Arg</td><td>Ala</td><td>A3P</td><td>Ala</td>

<td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>Ala</td><td>Pro Thr</td><td>Val</td><td>Ser</td><td>Ile</td><td>Phe</td><td>Pro</td><td>Pro</td><td>Ser Ser</td><td>Glu</td><td>Gin</td><td>Leu</td><td>Thr</td><td>Ser</td>

<td></td><td>115</td><td></td><td></td><td></td><td></td><td>120</td><td></td><td></td><td></td><td>125</td><td></td><td></td><td></td>

<td>Gly</td><td>Gly Ala</td><td>Ser</td><td>Val</td><td>Val</td><td>Cys</td><td>Phe</td><td>Leu</td><td>Asn Asn</td><td>Phe</td><td>Tyr</td><td>Pro</td><td>Lys</td><td>A3p</td>

<td></td><td>130</td><td></td><td></td><td></td><td>135</td><td></td><td></td><td></td><td>140</td><td></td><td></td><td></td><td></td>

<td>Ile</td><td>Asn Val</td><td>Lys</td><td>Trp</td><td>Lys</td><td>Ile</td><td>Asp</td><td>Gly</td><td>Ser Glu</td><td>Arg</td><td>Gin</td><td>Aen</td><td>Gly</td><td>Val</td>

<td>145</td><td></td><td></td><td></td><td>150</td><td></td><td></td><td></td><td>155</td><td></td><td></td><td></td><td></td><td>160</td>

<td>Leu</td><td>Asn Ser</td><td>Trp</td><td>Thr</td><td>Asp</td><td>Gin</td><td>Asp</td><td>Ser</td><td>Lvs Asp</td><td>Ser</td><td>Thr</td><td>Tyr</td><td>Ser</td><td>Met</td>

<td></td><td></td><td></td><td>165</td><td></td><td></td><td></td><td></td><td>170</td><td></td><td></td><td></td><td>175</td><td></td>

<td>Ser</td><td>Ser Thr</td><td>Leu</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Lys</td><td>Asp</td><td>Glu Tyr</td><td>Glu</td><td>Arg</td><td>His</td><td>A3n</td><td>Ser</td>

<td></td><td></td><td>180</td><td></td><td></td><td></td><td></td><td>185</td><td></td><td></td><td></td><td>190</td><td></td><td></td>

<td>Tyr</td><td>Thr Cys</td><td>Glu</td><td>Ala</td><td>Thr</td><td>His</td><td>Lys</td><td>Thr</td><td>Ser Thr</td><td>Ser</td><td>Pro</td><td>Ile</td><td>Val</td><td>Lys</td>

<td></td><td>195</td><td></td><td></td><td></td><td></td><td>200</td><td></td><td></td><td></td><td>205</td><td></td><td></td><td></td>

<td>Ser</td><td>Phe Asn</td><td>Arg</td><td>Asn</td><td>Glu</td><td>Cys</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td></td><td>210</td><td></td><td></td><td></td><td>215</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">&lt;210&gt; 177</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">&lt;211&gt; 1335</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="2">&lt;212&gt; DNA</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<td colspan="4">&lt;213&gt; Mus musculus</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td>

<400> 177

PZ/4572/AGR

141

EP 2 569 010 B1 gaggtccagc tgcaacaatc tggacctgag ctggtgaagc ctggggcttc agtgaagatg 60 tcctgtaagg cttctggata cacattcact gactactaca tgaactgggt gaagcagagc 120 catggagaga gccttgagtg gattggagat attaatcctt acaacgatga tactacctac 180 aaccacaagt tcaagggcaa ggccacattg actgtagaca aatcctccaa cacagcctac 240 atgcagctca acagcctgac atctgaggac tctgcagtct attactgtgc aagagagacg 300 gccgttatta ctacgaatgc tatggactac tggggtcaag gaacctcagt caccgtctcc 360 tcagccaaaa cgacaccccc atctgtctat ccactggccc ctggatctgc tgcccaaact 420 aactccatgg tgaccctggg atgcctggtc aagggctatt tccctgagcc agtgacagtg 480 acctggaact ctggatccct gtccagcggt gtgcacacct tcccagctgt cctgcagtct 540 gacctctaca ctctgagcag ctcagtgact gtcccctcca gcacctggcc cagcgagacc 600 gtcacctgca acgttgccca cccggccagc agcaccaagg tggacaagaa aattgtgccc 660 agggattgtg gttgtaagcc ttgcatatgt acagtcccag aagtatcatc tgtcttcatc 720 ttccccccaa agcccaagga tgtgctcacc attactctga ctcctaaggt cacgtgtgtt 780 gtggtagaca tcagcaagga tgatcccgag gtccagttca gctggtttgt agatgatgtg 840 gaggtgcaca cagctcagac gcaaccccgg gaggagcagt tcaacagcac tttccgctca 900 gtcagtgaac ttcccatcat gcaccaggac tggctcaatg gcaaggagtt caaatgcagg 960 gtcaacagtg cagctttccc tgcccccatc gagaaaacca tctccaaaac caaaggcaga 1020 ccgaaggctc cacaggtgta caccattcca cctcccaagg agcagatggc caaggataaa 1080 gtcagtctga cctgcatgat aacagacttc ttccctgaag acattactgt ggagtggcag 1140 tggaatgggc agccagcgga gaactacaag aacactcagc ccatcatgga cacagatggc 1200 tcttacttca tctacagcaa gctcaatgtg cagaagagca actgggaggc aggaaatact 1260 ttcacctgct ctgtgttaca tgagqqcctq cacaaccacc atactqaqaa gagcctctcc 1320 cactctcctg gtaaa 1335 <210> 178 <211> 445 <212> PRT <213> Mus musculus <400> 178

PZ/4572/AGR

142

EP 2 569 010 B1

Glu Val Gin Leu Gin Gin Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 15 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Tyr Met Asn Trp Val Lys Gin Ser His Gly Glu Ser Leu Glu Trp Ile 35 40 45

Gly Asp Ile Asn Pro Tyr Asn Asp Asp Thr Thr Tyr Asn His Lys Phe 50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80

Met Gin Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Glu Thr Ala Val Ile Thr Thr Asn Ala Met Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser 115 120 125

Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gin Thr Asn Ser Met Val 130 135 140

Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro

PZ/4572/AGR

143

EP 2 569 010 B1

180

185

190

Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro 195 200 205

Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly 210 215 220

Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile 225 230 235 240

Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys 245 250 255

Val Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gin 260 265 270

Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gin Thr Gin 275 280 285

Pro Arg Glu Glu Gin Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu 290 295 300

Pro Ile Met His Gin Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg 305 310 315 320

Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335

Thr Lys Gly Arg Pro Lys Ala Pro Gin Val Tyr Thr Ile Pro Pro Pro 340 345 350

Lys Glu Gin Met Ala Ly3 Asp Lys Val Ser Leu Thr Cys Met Ile Thr 355 360 365

Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gin Trp Asn Gly Gin 370 375 380

Pro Ala Glu Asn Tyr Lys Asn Thr Gin Pro Ile Met Asp Thr Asp Gly 385 390 395 400

Ser Tyr Phe Ile Tyr Ser Lys Leu Asn Val Gin Lys Ser Asn Trp Glu 405 410 415

Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn 420 425 430

His His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys 435 440 445

PZ/4572/AGR

144

EP 2 569 010 B1 <210> 179 <211> 324 <212> DNA <213> Mus musculus <400> 179

<td>gacatccagc</td><td>tgacccagag</td><td>ccccagcttc</td><td>ctttccgcat</td><td>ccgttggtga</td><td>ccgagtaaca</td><td>60</td>

<td>atcacatgcc</td><td>gcgcctcatc</td><td>ttcagttaca</td><td>tcttcttatc</td><td>ttaattggta</td><td>tcaacaaaaa</td><td>120</td>

<td>ccaggaaaag</td><td>cacctaaact</td><td>tcttatatac</td><td>tctacatcta</td><td>atctcgcatc</td><td>aggagttccc</td><td>180</td>

<td>tctcgatttt</td><td>caggatctgg</td><td>atcaggcaca</td><td>gaatttacac</td><td>ttactatatc</td><td>atcactccaa</td><td>240</td>

<td>ccagaagact</td><td>tcgccactta</td><td>ttactgccaa</td><td>caatacgatt</td><td>tttttccaag</td><td>cacattcgga</td><td>300</td>

<td>ggaggtacaa</td><td>aagtagaaat</td><td>caag</td><td></td><td></td><td></td><td>324</td>

<210> 180 <211> 108 <212> PRT <213> Mus musculus

<td colspan="16">&lt;400&gt; 180</td>

<td>Asp 1</td><td colspan="2">Ile Gin</td><td>Leu</td><td colspan="2">Thr Gin 5</td><td>Ser</td><td colspan="2">Pro Ser</td><td>Phe 10</td><td>Leu</td><td>Ser</td><td>Ala</td><td>Ser</td><td>Val 15</td><td>Gly</td>

<td>Aep</td><td>Arg</td><td>Val</td><td>Thr</td><td>Ile</td><td>Thr</td><td>Cys</td><td>Arg</td><td>Ala</td><td>Ser</td><td>Ser</td><td>Ser</td><td>Val</td><td>Thr</td><td>Ser</td><td>Ser</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>30</td><td></td><td></td>

<td>Tyr</td><td>Leu</td><td>Asn</td><td>Trp</td><td>Tyr</td><td>Gin</td><td>Gin</td><td>Lys</td><td>Pro</td><td>Gly</td><td>Lys</td><td>Ala</td><td>Pro</td><td>Lys</td><td>Leu</td><td>Leu</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Ile</td><td>Tyr</td><td>Ser</td><td>Thr</td><td>Ser</td><td>Asn</td><td>Leu</td><td>Ala</td><td>Ser</td><td>Gly</td><td>Val</td><td>Pro</td><td>Ser</td><td>Arg</td><td>Phe</td><td>Ser</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Gly</td><td>Ser</td><td>Gly</td><td>Ser</td><td>Gly</td><td>Thr</td><td>Glu</td><td>Phe</td><td>Thr</td><td>Leu</td><td>Thr</td><td>Ile</td><td>Ser</td><td>Ser</td><td>Leu</td><td>Gin</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Pro</td><td>Glu</td><td>Asp</td><td>Phe</td><td>Ala</td><td>Thr</td><td>Tyr</td><td>Tyr</td><td>Cys</td><td>Gin</td><td>Gin</td><td>Tyr</td><td>Asp</td><td>Phe</td><td>Phe</td><td>Pro</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>Ser</td><td>Thr</td><td>Phe</td><td>Gly</td><td>Gly</td><td>Gly</td><td>Thr</td><td>Ly3</td><td>Val</td><td>Glu</td><td>Ile</td><td>Lys</td><td></td><td></td><td></td><td></td>

100 105 <210> 181 <211> 363 <212> DNA <213> Mus musculus <400> 181

<td>gaggtgcagc</td><td>tggtgcagag</td><td>cggcgccgag</td><td>gtcaagaaac</td><td>ctggagcaag</td><td>cgtaaaggtt</td><td>60</td>

<td>agttgcaaag</td><td>catctggata</td><td>cacatttacc</td><td>gactactaca</td><td>tgaattgggt</td><td>acgacaagcc</td><td>120</td>

<td>cctggacaaa</td><td>gacttgaatg</td><td>gatgggagac</td><td>attaaccctt</td><td>ataacgacga</td><td>cactacatac</td><td>180</td>

<td>aatcataaat</td><td>ttaaaggaag</td><td>agttacaatt</td><td>acaagagata</td><td>catccgcatc</td><td>aaccgcctat</td><td>240</td>

<td>atggaacttt</td><td>cctcattgag</td><td>atctgaagac</td><td>actgctgttt</td><td>attactgtgc</td><td>aagagaaact</td><td>300</td>

<td>gccgttatta</td><td>ctactaacgc</td><td>tatggattac</td><td>tggggtcaag</td><td>gaaccactgt</td><td>taccgtctct</td><td>360</td>

<td>agt</td><td></td><td></td><td></td><td></td><td></td><td>363</td>

PZ/4572/AGR

145

EP 2 569 010 B1 <210> 182 <211> 121 <212> PRT <213> Mus musculus <400> 182

Glu Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30

Tyr Met Asn Trp Val Arg Gin Ala Pro Gly Gin Arg Leu Glu Trp Met 35 40 45

Gly Asp Ile Asn Pro Tyr Asn Asp Asp Thr Thr Tyr Asn His Lys Phe 50 55 60

Lys Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Glu Thr Ala Val Ile Thr Thr Asn Ala Met Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 183 <211> 363 <212> DNA <213> Sekwencja sztuczna <220>

<223> Sekwencja przeciwciała humanizowanego <400> 183

<td>gaggtgcagc</td><td>tggtgcagag</td><td>cggcgccgag</td><td>gtcaagaaac</td><td>ctggagcaag</td><td>cgtaaaggtt</td><td>60</td>

<td>agttgcaaag</td><td>catctggata</td><td>cacatttacc</td><td>gactactaca</td><td>tgaattgggt</td><td>acgacaagcc</td><td>120</td>

<td>cctggacaaa</td><td>gacttgaatg</td><td>gatgggagac</td><td>attaaccctt</td><td>ataacgacga</td><td>cactacatac</td><td>180</td>

<td>aatcataaat</td><td>ttaaaggaag</td><td>agttacaatt</td><td>acaagagata</td><td>catccgcatc</td><td>aaccgcctat</td><td>240</td>

<td>atggaacttt</td><td>cctcattgag</td><td>atctgaagac</td><td>actgctgttt</td><td>attactgtgc</td><td>aagagaaact</td><td>300</td>

<td>gccgttatta</td><td>ctactaacgc</td><td>tatggattac</td><td>tggggtcaag</td><td>gaaccactgt</td><td>taccgtctct</td><td>360</td>

agt 363 <210> 184 <211> 215 <212> PRT <213> Sekwencja sztuczna <220>

<223> Sekwencja przeciwciała humanizowanego <400> 184

PZ/4572/AGR

146

EP 2 569 010 B1

Asp Ile Gin Leu Thr 1 5

Gin Ser Pro Ser Phe 10

Leu Ser Ala Ser Val Gly 15

Asp Arg Val Thr Ile 20

Thr Cys Arg Ala Ser 25

Ser Ser Val Thr Ser Ser 30

Tyr Leu Asn Trp Tyr 35

Gin Gin Lys Pro Gly 40

Lys Ala Pro Lys Leu Leu 45

Ile Tyr Ser Thr Ser 50

Asn Leu Ala Ser Gly 55

Val Pro Ser Arg Phe Ser 60

Gly Ser Gly Ser Gly 65

Thr Glu Phe Thr Leu 70

Thr Ile Ser Ser Leu Gin 75 80

Pro Glu Asp Phe Ala 85

Thr Tyr Tyr Cys Gin 90

Gin Tyr Asp Phe Phe Pro 95

Ser Thr Phe Gly Gly 100

Gly Thr Lys Val Glu 105

Ile Lys Arg Thr Val Ala 110

Ala Pro Ser Val Phe 115

Ile Phe Pro Pro Ser 120

Asp Glu Gin Leu Lys Ser 125

Gly Thr Ala Ser Val 130

Val Cys Leu Leu Asn 135

Asn Phe Tyr Pro Arg Glu 140

Ala Lys Val Gin Trp 145

Lys Val Asp Asn Ala 150

Leu Gin Ser Gly Asn Ser 155 160

Gin Glu Ser Val Thr 165

Glu Gin Asp Ser Lys 170

Asp Ser Thr Tyr Ser Leu 175

Ser Ser Thr Leu Thr 180

Leu Ser Lys Ala Asp 185

Tyr Glu Lys His Lys Val 190

Tyr Ala Cys Glu Val 195

Thr His Gin Gly Leu 200

Ser Ser Pro Val Thr Lys 205

Ser Phe Asn Arg Gly 210 <210> 185 <211> 1341 <212> DNA

Glu Cys 215 <213> Sekwencja sztuczna <220>

<223> Sekwencja przeciwciała humanizowanego <400> 185

PZ/4572/AGR

147

EP 2 569 010 B1

<td>gaggtgcagc</td><td>tggtgcagag</td><td>cggcgccgag</td><td>gtcaagaaac</td><td>ctggagcaag</td><td>cgtaaaggtt</td><td>60</td>

<td>agttgcaaag</td><td>catctggata</td><td>cacatttacc</td><td>gactactaca</td><td>tgaattgggt</td><td>acgacaagcc</td><td>120</td>

<td>cctggacaaa</td><td>gacttgaatg</td><td>gatgggagac</td><td>attaaccctt</td><td>ataacgacga</td><td>cactacatac</td><td>180</td>

<td>aatcataaat</td><td>ttaaaggaag</td><td>agttacaatt</td><td>acaagagata</td><td>catccgcatc</td><td>aaccgcctat</td><td>240</td>

<td>atggaacttt</td><td>cctcattgag</td><td>atctgaagac</td><td>actgctgttt</td><td>attactgtgc</td><td>aagagaaact</td><td>300</td>

<td>gccgttatta</td><td>ctactaacgc</td><td>tatggattac</td><td>tggggtcaag</td><td>gaaccactgt</td><td>taccgtctct</td><td>360</td>

<td>agtgcctcca</td><td>ccaagggccc</td><td>atcggtcttc</td><td>cccctggcgc</td><td>cctgctccag</td><td>gagcacctcc</td><td>420</td>

<td>gagagcacag</td><td>cggccctggg</td><td>ctgcctggtc</td><td>aaggactact</td><td>tccccgaacc</td><td>ggtgacggtg</td><td>480</td>

<td>tcgtggaact</td><td>caggcgctct</td><td>gaccagcggc</td><td>gtgcacacct</td><td>tcccagctgt</td><td>cctacagtcc</td><td>540</td>

<td>tcaggactct</td><td>actccctcag</td><td>cagcgtggtg</td><td>accgtgccct</td><td>ccagcaactt</td><td>cggcacccag</td><td>600</td>

<td>acctacacct</td><td>gcaacgtaga</td><td>tcacaagccc</td><td>agcaacacca</td><td>aggtggacaa</td><td>gacagttgag</td><td>660</td>

<td>cgcaaatgtt</td><td>gtgtcgagtg</td><td>cccaccgtgc</td><td>ccagcaccac</td><td>ctgtggcagg</td><td>accgtcagtc</td><td>720</td>

<td>ttcctcttcc</td><td>ccccaaaacc</td><td>caaggacacc</td><td>ctcatgatct</td><td>cccggacccc</td><td>tgaggtcacg</td><td>780</td>

<td>tgcgtggtgg</td><td>tggacgtgag</td><td>ccacgaagac</td><td>cccgaggtcc</td><td>agttcaactg</td><td>gtacgtggac</td><td>840</td>

<td>ggcgtggagg</td><td>tgcataatgc</td><td>caagacaaag</td><td>ccacgggagg</td><td>agcagttcaa</td><td>cagcacgttc</td><td>900</td>

<td>cgtgtggtca</td><td>gcgtcctcac</td><td>cgttgtgcac</td><td>caggactggc</td><td>tgaacggcaa</td><td>ggagtacaag</td><td>960</td>

<td>tgcaaggtct</td><td>ccaacaaagg</td><td>cctcccagcc</td><td>cccatcgaga</td><td>aaaccatctc</td><td>caaaaccaaa</td><td>1020</td>

<td>gggcagcccc</td><td>gagaaccaca</td><td>ggtgtacacc</td><td>ctgcccccat</td><td>cccgggagga</td><td>gatgaccaag</td><td>1080</td>

<td>aaccaggtca</td><td>gcctgacctg</td><td>cctggtcaaa</td><td>ggcttctacc</td><td>ccagcgacat</td><td>cgccgtggag</td><td>1140</td>

<td>tgggagagca</td><td>atgggcagcc</td><td>ggagaacaac</td><td>tacaagacca</td><td>cacctcccat</td><td>gctggactcc</td><td>1200</td>

<td>gacggctcct</td><td>tcttcctcta</td><td>cagcaagctc</td><td>accgtggaca</td><td>agagcaggtg</td><td>gcagcagggg</td><td>1260</td>

<td>aacgtcttct</td><td>catgctccgt</td><td>gatgcatgag</td><td>gctctgcaca</td><td>accactacac</td><td>gcagaagagc</td><td>1320</td>

<td>ctctccctgt</td><td>ctccgggtaa</td><td>a</td><td></td><td></td><td></td><td>1341</td>

<210> 186 <211> 447 <212> PRT <213> Sekwencja sztuczna <220>

<223> Sekwencja przeciwciała humanizowanego <400> 186

EP 2 569 010 B1

<td colspan="9">PZ/4572/AGR</td><td colspan="2" rowspan="2">148 Glu Val 10</td><td rowspan="2">Lys</td><td rowspan="2">Lys</td><td rowspan="2">Pro</td><td rowspan="2">Gly 15</td><td rowspan="2">Ala</td>

<td>Glu 1</td><td>Val</td><td>Gin</td><td>Leu</td><td colspan="2">Val Gin 5</td><td>Ser</td><td colspan="2">Gly Ala</td>

<td>Ser</td><td>Val</td><td>Lys</td><td>Val</td><td>Ser</td><td>Cys</td><td>Lys</td><td>Ala</td><td>Ser</td><td>Gly</td><td>Tyr</td><td>Thr</td><td>Phe</td><td>Thr</td><td>Asp</td><td>Tyr</td>

<td></td><td></td><td></td><td>20</td><td></td><td></td><td></td><td></td><td>25</td><td></td><td></td><td></td><td></td><td>30</td><td></td><td></td>

<td>Tyr</td><td>Met</td><td>Asn</td><td>Trp</td><td>Val</td><td>Arg</td><td>Gin</td><td>Ala</td><td>Pro</td><td>Gly</td><td>Gin</td><td>Arg</td><td>Leu</td><td>Glu</td><td>Trp</td><td>Met</td>

<td></td><td></td><td>35</td><td></td><td></td><td></td><td></td><td>40</td><td></td><td></td><td></td><td></td><td>45</td><td></td><td></td><td></td>

<td>Gly</td><td>Asp</td><td>Ile</td><td>Asn</td><td>Pro</td><td>Tyr</td><td>Asn</td><td>Asp</td><td>Asp</td><td>Thr</td><td>Thr</td><td>Tyr</td><td>Asn</td><td>His</td><td>Lys</td><td>Phe</td>

<td></td><td>50</td><td></td><td></td><td></td><td></td><td>55</td><td></td><td></td><td></td><td></td><td>60</td><td></td><td></td><td></td><td></td>

<td>Lys</td><td>Gly</td><td>Arg</td><td>Val</td><td>Thr</td><td>Ile</td><td>Thr</td><td>Arg</td><td>Asp</td><td>Thr</td><td>Ser</td><td>Ala</td><td>Ser</td><td>Thr</td><td>Ala</td><td>Tyr</td>

<td>65</td><td></td><td></td><td></td><td></td><td>70</td><td></td><td></td><td></td><td></td><td>75</td><td></td><td></td><td></td><td></td><td>80</td>

<td>Met</td><td>Glu</td><td>Leu</td><td>Ser</td><td>Ser</td><td>Leu</td><td>Arg</td><td>Ser</td><td>Glu</td><td>Asp</td><td>Thr</td><td>Ala</td><td>Val</td><td>Tyr</td><td>Tyr</td><td>Cys</td>

<td></td><td></td><td></td><td></td><td>85</td><td></td><td></td><td></td><td></td><td>90</td><td></td><td></td><td></td><td></td><td>95</td><td></td>

<td>Ala</td><td>Arg</td><td>Glu</td><td>Thr</td><td>Ala</td><td>Val</td><td>Ile</td><td>Thr</td><td>Thr</td><td>Asn</td><td>Ala</td><td>Met</td><td>Asp</td><td>Tyr</td><td>Trp</td><td>Gly</td>

<td></td><td></td><td></td><td>100</td><td></td><td></td><td></td><td></td><td>105</td><td></td><td></td><td></td><td></td><td>110</td><td></td><td></td>

<td>Gin</td><td>Gly</td><td>Thr</td><td>Thr</td><td>Val</td><td>Thr</td><td>Val</td><td>Ser</td><td>Ser</td><td>Ala</td><td>Ser</td><td>Thr</td><td>Lys</td><td>Gly</td><td>Pro</td><td>Ser</td>

PZ/4572/AGR

149

EP 2 569 010 B1

115 120 125

Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Asn Phe Gly Thr Gin Thr Tyr Thr Cys Asn Val Asp His 195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys 210 215 220

Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val 225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270

Val Gin Phe Asn Trp Tyr Val Asp Gly Val Glu Val Hi3 Asn Ala Lye 275 280 285