Binding proteins specific for insulin-like growth factors and uses thereof in the preparation of medicaments for the treatment of malignant tumors

191718/3

BINDING PROTEINS SPECIFIC FOR INSULIN-LIKE GROWTH FACTORSAND USES THEREOF IN THE PREPARATION OF MEDICAMENTS FOR THETREATMENT OF MALIGNANT TUMORS

BACKGROUND OF THE INVENTION

Field of the Invention [0002] The invention relates to binding proteins that bind to insulin-like growthfactor-2 (IGF-II) with cross-reactivity to insulin-like growth factor-1 (IGF-I) and uses ofsuch binding proteins. More specifically, the invention relates to monoclonal antibodiesdirected to IGF-II with cross-reactivity to IGF-I and uses of these antibodies. Aspects ofthe invention also relate to hybridomas or other cell lines expressing such antibodies.

Description of the Related Art [0003] Insulin-like growth factor IGF-I and IGF-II are small polypeptidesinvolved in regulating cell proliferation, survival, differentiation and transformation. IGFsexert their various actions by primarily interacting with a specific cell surface receptor, theIGF-I receptor (IGF-IR) and activating various intracellular signaling cascades. IGFscirculate in serum mostly bound to IGF-binding proteins (IGFBP-I to 6). The interaction ofIGFs with the IGF-IR is regulated by the IGFBPs, and IGFs can only bind to the IGF-IRonce released from the IGFBPs (mostly by proteolysis of the IGFBPs). IGF-I can also bindto a hybrid receptor comprised of IGF-IR and insulin receptor (IR) subunits. IGF-II hasbeen shown to bind to the "A" isoform of the insulin receptor.

[0004] Malignant transformation involves the imbalance of diverse processessuch as cell growth, differentiation, apoptosis, and transformation. IGF-I and IGF-II have 1 P/15804/131320/608077/1 WO 2007/070432 PCT/US2006/047059 been implicated in the pathophysiology of a wide range of conditions, and are thought toplay a role in tumorigenesis due to the mitogenic and antiapoptotic properties mediatedby the receptor IGF-IR. LeRoith and Roberts, Cancer Lett. 195:127-137 (2003).

[0005] IGF-I was discovered as a growth factor produced by the liver underthe regulatory control of pituitary growth hormone and was originally designatedsomatomedin-C. Salmon et al., J. Lab. Clin. Med. 49:825-826 (1957). Both IGF-I andIGF-II are expressed ubiquitously and act as endocrine, paracrine, and autocrine growthfactors, through their interaction with the IGF-IR, a trans-membrane tyrosine kinase thatis structurally and functionally related to the insulin receptor (IR). IGF-I functionsprimarily by activating the IGF-IR, whereas IGF-Π can act through either the IGF-IR orthrough the IR-A isoform. LeRoith and Roberts, Cancer Lett. 195:127-137 (2003).Additionally, the interaction of both IGF-I and IGF-II with the IGF-binding proteins mayaffect the half-life and bioavailability of the IGFs, as well as their direct interaction withreceptors in some cases. Rajaram et al., Endocr. Rev. 18:801-831 (1997).

[0006] IGF-I has a long-term impact on cell proliferation, differentiation, andapoptosis. Experiments in cultured osteosarcoma and breast cancer cells suggested thatIGF-I is a potent mitogen and exerts its mitogenic action by increasing DNA synthesisand by stimulating the expression of cyclin DI, which accelerates progression of the cellcycle from Gi to S phase. Furlanetto et al., Mol. Endocrinol. 8:510-517 (1994); Dufoumyet al., J. Biol. Chem. 272:311663-31171 (1997). Suppression of cyclin DI expression inpancreatic cancer cells abolished the mitogenic effect of IGF-I. Kornmann et al., J. Clin.Invest. 101:344-352 (1998). In addition to stimulating cell cycle progression, IGF-I alsoinhibits apoptosis. IGF-I was shown to stimulate the expression of Bel proteins and tosuppress expression of Bax, which results in an increase in the relative amount of theBcl/Bax heterodimer, thereby blocking initiation of the apoptotic pathway. Minshall etal., J. Immunol. 159:1225-1232 (1997); Parrizas et al., Endocrinology 138:1355-1358(1997); Wang et al., Endocrinology 139:1354-1360 (1998).

[0007] Like IGF-I, IGF-II also has mitogenic and antiapoptotic actions andregulates cell proliferation and differentiation. Compared with IGF-I, high concentrationsof IGF-II circulate in serum. High serum IGF-Π concentrations have been found inpatients with colorectal cancer, with a trend towards higher concentrations in advanceddisease. Renehan et al., Br. J. Cancer 83:1344-1350. Additionally, most primary tumorsand transformed cell lines overexpress IGF-II mRNA and protein. Werner and LeRoithAdv. Cancer Res. 68:183-223 (1996). Overexpression of IGF-Π in colon cancer is 2 WO 2007/070432 PCT/US2006/047059 associated with an aggressive phenotype, and the loss of imprinting (loss of allele-specificexpression) of the IGF-II gene may be important in colorectal carcinogenesis. Michell etal., Br. J. Cancer 76:60-66 (1997); Takano et al., Oncology 59:210-216 (2000). Cancercells with a strong tendency to metastasize have four-fold higher levels of IGF-IIexpression than those cells with a low ability to metastasize. Guerra et al., Int. J. Cancer65:812-820(1996).

[0008] Research and clinical studies have highlighted the role of the IGFfamily members in the development, maintenance and progression of cancer. Manycancer cells have been shown to overexpress the IGF-IR and/or the IGF ligands. Forexample, IGF-I and IGF-II are strong mitogens for a wide variety of cancer cell lines,including sarcoma, leukemia, and cancers of the prostate, breast, lung, colon, stomach,esophagus, liver, pancreas, kidney, thyroid, brain, ovary, and uterus. Macaulay el al., Br.J. Cancer 65:311-320 (1992); Oku et al., Anticancer Res. 11:1591-1595 (1991); LeRoithet al., Ann. Intern. Med. 122:54-59 (1995); Yaginuma et al., Oncology 54:502-507(1997); Singh et al., Endocrinology 137:1764-1774 (1996); Frostad et al., Eur. J.Haematol 62:191-198 (1999). When IGF-I was administered to malignant colon cancercells, they became resistant to cytokine-induced apoptosis. Remacle-Bonnet et al.,Cancer Res. 60:2007-2017 (2000).

[0009] The role of IGFs in cancer is also supported by epidemiologic studies,which showed that high levels of circulating IGF-I and low levels of IGFBP-3 areassociated with an increased risk for development of several common cancers (prostate,breast, colorectal and lung). Mantzoros et al., Br. J. Cancer 76:1115-1118 (1997);Hankinson et al., Lancet 351:1393-1396 (1998); Ma et al., J. NatLCancer Inst. 91:620-625 (1999); Karasik et al., J. Clin. Endocrinol Metab. 78:271-276 (1994). These resultssuggest that IGF-I and IGF-II act as powerful mitogenic and anti-apoptotic signals, andthat their overexpression correlates with poor prognosis in patients with several types ofcancer.

[0010] Using knockout mouse models, several studies have further establishedthe role of IGFs in tumor growth. With the development of the technology for tissuespecific, conditioned gene deletion, a mouse model of liver IGF-I deficiency (LID) wasdeveloped. Liver-specific deletion of the igfl gene abrogated expression of IGF-I mRNAand caused a dramatic reduction in circulating IGF-I levels. Yakar et al., Proc. Natl.Acad. Sci. USA 96:7324-7329 (1999). When mammary tumors were induced in the LIDmouse, reduced circulating IGF-1 levels resulted in significant reductions in cancer 3 WO 2007/070432 PCT/US2006/047059 development, growth, and metastases, whereas increased circulating IGF-1 levels were associated with enhanced tumor growth. Wu et al., Cancer Res. 63:4384-4388 (2003).

[0011] Several papers have reported that inhibition of IGF-IR expressionand/or signaling leads to inhibition of tumor growth, both in vitro and in vivo. Inhibitionof IGF signaling has also been shown to increase the susceptibility of tumor cells tochemotherapeutic agents. A variety of strategies (antisense oligonucleotides, solublereceptor, inhibitory peptides, dominant negative receptor mutants, small moleculesinhibiting the kinase activity and anti-hIGF-IR antibodies) have been developed to inhibitthe IGF-IR signaling pathway in tumor cells. One approach has been to target the kinaseactivity of IGF-IR with small molecule inhibitors. Two compounds were recentlyidentified as small molecule kinase inhibitors capable of selectively inhibiting the IGF-IR.Garcia-Echeverria et al., Cancer Cell 5:231-239 (2004); Mitsiades et al., Cancer Cell5:221-230 (2004). Inhibition of IGF-IR kinase activity abrogated IGF-I-mediatedsurvival and colony formation in soft agar of MCF-7 human breast cancer cells. Garcia-Echeverria et al., Cancer Cell 5:231-239 (2004). When an IGF-IR kinase inhibitor wasadministered to mice bearing tumor xenografts, IGF-IR signaling in tumor xenografts wasinhibited and the growth of IGF-IR-driven fibrosarcomas was significantly reduced.Garcia-Echeverria et al., Cancer Cell 5:231-239 (2004). A similar effect was observedon hematologic malignancies, especially multiple myeloma. In multiple myeloma cells, asmall molecule IGF-IR kinase inhibitor demonstrated a > 16-fold greater potency againstthe IGF-IR, as compared to the insulin receptor, and was similarly effective in inhibitingcell growth and survival. Mitsiades et al., Cancer Cell 5:221-230 (2004). The samecompound was injected intraperitoneally into mice and inhibited multiple myeloma cellgrowth and enhanced survival of the mice. Mitsiades et al., Cancer Cell 5:221-230(2004). When combined with other chemotherapeutics at subtherapeutic doses, inhibitionof IGF-IR kinase activity synergistically reduced tumor burden. Mitsiades et al., CancerCell 5:221-230 (2004).

[0012] Another approach to inhibit IGF signaling has been the development ofneutralizing antibodies directed against the receptor IGF-IR. Various groups havedeveloped antibodies to IGF-IR that inhibit receptor IGF-I-stimulatedautophosphorylation, induce receptor internalization and degradation, and reduceproliferation and survival of diverse human cancer cell lines. Hailey et al., Mol CancerTher. 1:1349-1353 (2002); Maloney et al., Cancer Res. 63:5073-5083 (2003); Benini etal., Clin. Cancer Res. 7:1790-1797 (2001); Burtrum et al., Cancer Res. 63:8912-8921 4 WO 2007/070432 PCT/US2006/047059 (2003). Additionally, in xenograft tumor models, IGF-IR blockade resulted in significantgrowth inhibition of breast, renal and pancreatic tumors in vivo. Burtrum et al., CancerRes. 63:8912-8921 (2003); Maloney et al., Cancer Res. 63:5073-5083 (2003).Experiments utilizing chimeric humanized IGF-IR antibodies yielded similar results,inhibiting growth of breast cancer cells in vitro and in tumor xenografts. Sachdev et al.,Cancer Res. 63:627-635 (2003). Other humanized IGF-IR antibodies blocked IGF-I-induced tyrosine phosphorylation and growth inhibition in breast and non small cell lungtumors, as well as in vivo. Cohen et al., Clin. Cancer Res. 11:2063-2073 (2005); Goetschetal., Int. J. Cancer 113:316-328 (2005).

[0013] Increased IGF-I levels have also been associated with several non-cancerous pathological conditions, including acromegaly and gigantism (Barkan,Cleveland Clin. J. Med. 65: 343, 347-349, 1998), while abnormal IGF-I/IGF-II receptorfunction has been implicated in psoriasis (Wraight et al., Nat. Biotech. 18: 521-526,2000), atherosclerosis and smooth muscle restenosis of blood vessels followingangioplasty (Bayes-Genis et al., Circ. Res. 86: 125-130, 2000). Increased IGF-I levelshave been implicated in diabetes or in complications associated with diabetes, such asmicrovascular proliferation (Smith et al., Nat. Med. 5: 1390-1395, 1999).

[0014] Antibodies to IGF-I and IGF-II have been disclosed in the art. See, forexample, Goya et al., Cancer Res. 64:6252-6258 (2004); Miyamoto et al., Clin. CancerRes. 11:3494-3502 (2005). Additionally, see WO 05/18671, WO 05/28515 and WO03/93317.

SUMMARY

[0015] Embodiments of the invention relate to binding proteins thatspecifically bind to insulin-like growth factors and reduce tumor growth. In oneembodiment, the binding proteins are fully human monoclonal antibodies, or bindingfragments thereof that specifically bind to insulin-like growth factors and reduce tumorgrowth. Mechanisms by. which this can be achieved can include and are not limited toeither inhibition of binding of IGF-I/II to its receptor IGF-IR, inhibition of IGF-I/II-induced IGF-IR signaling, or increased clearance of IGF-I/II, therein reducing theeffective concentration of IGF-I/II.

[0016] Thus, some embodiments provide a fully human isolated specificbinding protein that preferentially binds to insulin-like growth factor-II (IGF-II) withcross-reactivity to insulin-like growth factor I (IGF-I) and neutralizes IGF-I and IGF-II 5 WO 2007/070432 PCT/US2006/047059 activity. In certain aspects, the binding protein binds to IGF-II with at least 2.5 timesgreater affinity than to IGF-I. In other aspects, the binding protein binds to IGF-II with atleast 3, at least 4, at least 5, at least 7, at least 10, at least 50, at least 60, at least 100 or atleast 150 times greater affinity than to IGF-I.

[0017} In some embodiments, the specific binding protein has an EC50 of nomore than 15 nM for inhibiting IGF-I-dependent IGF-I receptor phosphorylation inNIH3T3 cells expressing IGF-IR ectopically. In some aspects, the specific bindingprotein has an EC50 of no more than 15 nM, no more than 10 nM, or no more than 8 nMfor inhibiting IGF-I-dependent IGF-I receptor phosphorylation in NIH3T3 cellsexpressing IGF-IR ectopically.

[0018] In some embodiments, the specific binding protein has an EC50 of nomore than 5 nM, no more than 4 nM, or no more than 3 nM for inhibiting 1GF-II-dependent IGF-I receptor phosphorylation in NIH3T3 cells expressing IGF-IRectopically.

[0019] In other embodiments, the specific binding protein inhibits greater than70% of IGF-II dependent proliferation of NIH3T3 cells that express recombinant hlGF-IR with an EC50 of no more than 25 nM, no more than 20 nM, no more than 15 nM, or nomore than 10 nM.

[0020] In other embodiments, the specific binding protein inhibits greater than70% of IGF-I dependent proliferation of NIH3T3 cells that express recombinant hIGF-IRwith an EC50 of no more than 40 nM, no more than 30 nM, or no more than 25 nM.

[0021] In certain embodiments, the specific binding protein competes forbinding with a monoclonal antibody comprising a variable heavy chain sequence selectedfrom the group consisting of SEQ ID NO.: 2, SEQ ID NO.: 6, SEQ ID NO.: 10, SEQ IDNO.: 14 and SEQ ID NO.: 18, and comprising a variable light chain sequence selectedfrom the group consisting of SEQ ID NO.: 4, SEQ ID NO.: 8, SEQ ID NO.: 12 and SEQID NO.: 16.

[0022] One embodiment of the invention is a fully human antibody that bindsto IGF-I with a Kd less than 500 picomolar (pM). More preferably, the antibody bindswith a Kd less than 450 picomolar (pM). More preferably, the antibody binds with a Kdless than 410 picomolar (pM). More preferably, the antibody binds with a K<j of less than350 pM. Even more preferably, the antibody binds with a Kd of less than 300 pM.Affinity and/or avidity measurements can be measured by BIACORE®, as describedherein. 6 WO 2007/070432 PCT/US2006/047059 [0023] Yet another embodiment of the invention is a fully human monoclonalantibody that binds to IGF-II with a Ka of less than 175 picomolar (pM). Morepreferably, the antibody binds with a Kd less than 100 picomolar (pM). More preferably,the antibody binds with a Kd less than 50 picomolar (pM). More preferably, the antibodybinds with a Kd less than 5 picomolar (pM). Even more preferably, the antibody bindswith a Ka of less than 2 pM.

[0024] In certain embodiments, the specific binding protein is a fully humanmonoclonal antibody or a binding fragment of a fully human monoclonal antibody. Thebinding fragments can include fragments such as Fab, Fab’ or F(ab’)2 and Fv.

[0025] One embodiment of the invention comprises fully human monoclonalantibodies 7.251.3 (ATCC Accession Number PTA-7422), 7.34.1 (ATCC AccessionNumber PTA-7423) and 7.159.2 (ATCC Accession Number PTA-7424) whichspecifically bind to IGF-I/II, as discussed in more detail below.

[0026] In some embodiments the specific binding protein that binds to insulin-like growth factor-II (IGF-Π) with cross-reactivity to insulin-like growth factor-I (IGF-I),or binding fragment thereof can include a heavy chain polypeptide having the sequence ofSEQ ID NO.: 6, and a light chain polypeptide having the sequence of SEQ ID NO.: 8.

[0027] The specific binding protein can include a heavy chain polypeptidehaving the sequence of SEQ ID NO.: 10, and a light chain polypeptide having thesequence of SEQ ID NO.: 12.

[0028] The specific binding protein of the invention can include heavy chainpolypeptide having the sequence of SEQ ID NO.: 14 and a light chain polypeptide havingthe sequence of SEQ ID NO.: 16.

[0029] In certain embodiments, the specific binding protein can be in amixture with a pharmaceutically acceptable carrier.

[0030] Another embodiment includes isolated nucleic acid moleculesencoding any of the specific binding proteins described herein, vectors having isolatednucleic acid molecules encoding the specific binding proteins, or a host cell transformedwith any of such nucleic acid molecules and vectors.

[0031] In certain embodiments the specific binding protein that binds toinsulin-like growth factor-II (IGF-II) with cross-reactivity to insulin-like growth factor-I(IGF-I), or binding fragment thereof does not bind specifically to IGF-II or IGF-I proteinswhen said proteins are bound to Insulin Growth Factor Binding Proteins. 7 WO 2007/070432 PCT/US2006/047059 [0032] Further embodiments include methods of determining the level ofinsulin-like growth factor-Π (IGF-II) and insulin-like growth factor I (IGF-I) in a patientsample. These methods can include providing a patient sample; contacting the samplewith a specific binding protein that binds to insulin-like growth factor-II (IGF-II) withcross-reactivity to insulin-like growth factor-I (IGF-I), or binding fragment thereof; anddetermining the level of IGF-I and IGF-II in said sample. In some aspects, the patientsample is blood.

[0033] Additional embodiments include methods of treating a malignanttumor in a mammal. These methods can include selecting a mammal in need of treatmentfor a malignant tumor; and administering to the mammal a therapeutically effective doseof a specific binding protein that binds to insulin-like growth factor-II (IGF-II) withcross-reactivity to insulin-like growth factor-I (IGF-I), or binding fragment thereof. Insome aspects the animal is human. In some aspects the binding protein is a fully humanmonoclonal antibody, and is selected from the group consisting of mAb 7.251.3 (ATCCAccession Number PTA-7422), mAb 7.34.1 (ATCC Accession Number PTA-7423), andmAb 7.159.2 (ATCC Accession Number PTA-7424).

[0034] Treatable diseases can include melanoma, non-small cell lung cancer,glioma, hepatocellular (liver) carcinoma, thyroid tumor, gastric (stomach) cancer,prostrate cancer, breast cancer, ovarian cancer, bladder cancer, lung cancer, glioblastoma,endometrial cancer, kidney cancer, colon cancer, pancreatic cancer, and epidermoidcarcinoma.

[0035] Additional embodiments include methods of treating a growth factor-dependent disease in a mammal. These methods include selecting a mammal in need oftreatment for a growth factor-dependent disease; and administering to said mammal atherapeutically effective dose of a specific binding protein that binds to insulin-likegrowth factor-II (IGF-II) with cross-reactivity to insulin-like growth factor-I (IGF-I), orbinding fragment thereof. In some aspects, the mammal can be human. In some aspectsthe binding protein is a fully human monoclonal antibody, and is selected from the groupconsisting of mAb 7.251.3 (ATCC Accession Number PTA-7422), mAb 7.34.1 (ATCCAccession Number PTA-7423), and mAb 7.159.2 (ATCC Accession Number PTA-7424). (0036] Treatable growth factor-dependent diseases can include osteoporosis,diabetes, and cardiovascular diseases. Other treatable disease conditions includeacromegaly and gigantism, psoriasis, atherosclerosis and smooth muscle restenosis ofblood vessels, as well as diabetes. 8 WO 2007/070432 PCT/US2006/047059 [0037] Additional embodiments include a conjugate comprising a fully humanmonoclonal antibody that binds to insulin-like growth factor-II (IGF-II) with cross-reactivity to insulin-like growth factor-I (IGF-I), or a binding fragment thereof and atherapeutic agent. In some aspects the therapeutic agent can be a toxin, a radioisotope, ora pharmaceutical composition.

[0038] In other embodiments, the invention provides fully human monoclonalantibodies, or binding fragment thereof, that bind to insulin-like growth factor-II (IGF-II)with cross-reactivity to insulin-like growth factor-I (IGF-I), and comprise a heavy chaincomplementarity determining region 1 (CDR1) having the amino acid sequence of “SerTyr Tyr Trp Ser” (SEQ ID NO: 21); a heavy chain complementarity determining region 2(CDR2) having the amino acid sequence of “Tyr Phe Phe Tyr Ser Gly Tyr Thr Asn TyrAsn Pro Ser Leu Lys Ser” (SEQ ID NO: 22); and a heavy chain complementaritydetermining region 3 (CDR3) having the amino acid sequence of “lie Thr Gly Thr ThrLys Gly Gly Met Asp Val” (SEQ ID NO: 23).

[0039] Further embodiments include fully human monoclonal antibodies, orbinding fragment thereof, having a light chain complementarity determining region 1(CDR1) having the amino acid sequence of “Thr Gly Ser Ser Ser Asn lie Gly Ala Gly TyrAsp Val His” (SEQ ID NO: 24). Antibodies herein can also include a light chaincomplementarity determining region 2 (CDR2) having the amino acid sequence of “GlyAsn Asn Asn Arg Pro Ser” (SEQ ID NO: 25); and a light chain complementaritydetermining region 3 (CDR3) having the amino acid sequence of “Gin Ser Phe Asp SerSer Leu Ser Gly Ser Val” (SEQ ID NO: 26).

[0040] In other embodiments, the invention provides fully human monoclonalantibodies, or binding fragment thereof, that bind to insulin-like growth factor-Π (IGF-II)with cross-reactivity to insulin-like growth factor-I (IGF-I), and comprise a heavy chaincomplementarity determining region 1 (CDR1) having the amino acid sequence of “SerTyr Tyr Trp Ser” (SEQ ID NO: 27); a heavy chain complementarity determining region 2(CDR2) having the amino acid sequence of “Tyr Phe Phe Tyr Ser Gly Tyr Thr Asn TyrAsn Pro Ser Leu Lys Ser” (SEQ ID NO: 28); and a heavy chain complementaritydetermining region 3 (CDR3) having the amino acid sequence of “He Thr Gly Thr ThrLys Gly Gly Met Asp Val” (SEQ ID NO: 29).

[0041] Further embodiments include fully human monoclonal antibodies, orbinding fragment thereof, having a light chain complementarity determining region 1(CDR1) having the amino acid sequence of “Thr Gly Arg Ser Ser Asn lie Gly Ala Gly 9 WO 2007/070432 PCT/US2006/047059

Tyr Asp Val His” (SEQ ID NO: 30); a light chain complementarity determining region 2(CDR2) having the amino acid sequence of “Gly Asn Ser Asn Arg Pro Ser” (SEQ ID NO:31); and a light chain complementarity determining region 3 (CDR3) having the aminoacid sequence of “Gin Ser Tyr Asp Ser Ser Leu Ser Gly Ser Val” (SEQ ID NO: 32).

[0042] In other embodiments, the invention provides fully human monoclonalantibodies, or binding fragment thereof, that bind to insulin-like growth factor-II (IGF-II)with cross-reactivity to insulin-like growth factor-I (IGF-I), and comprise a heavy chaincomplementarity determining region 1 (CDR1) having the amino acid sequence of “SerTyr Asp lie Asn” (SEQ ID NO: 33); a heavy chain complementarity determining region 2(CDR2) having the amino acid sequence of “Trp Met Asn Pro Asn Ser Gly Asn Thr GlyTyr Ala Gin Lys Phe Gin Gly” (SEQ ID NO: 34); and a heavy chain complementaritydetermining region 3 (CDR3) having the amino acid sequence of “Asp Pro Tyr Tyr TyrTyr Tyr Gly Met Asp Val” (SEQ ID NO: 35).

[0043] Further embodiments include fully human monoclonal antibodies, orbinding fragment thereof, having a light chain complementarity determining region 1(CDR1) having the amino acid sequence of “Ser Gly Ser Ser Ser Asn lie Glu Asn AsnHis Val Ser” (SEQ ID NO: 36); a light chain complementarity determining region 2(CDR2) having the amino acid sequence of “Asp Asn Asn Lys Arg Pro Ser” (SEQ IDNO: 37); and a light chain complementarity determining region 3 (CDR3) having theamino acid sequence of “Glu Thr Trp Asp Thr Ser Leu Ser Ala Gly Arg Val” (SEQ IDNO: 38).

[0044] In other embodiments, the invention provides fully human monoclonalantibodies, or binding fragment thereof, that bind to insulin-like growth factor-II (IGF-II)with cross-reactivity to insulin-like growth factor-I (IGF-I), and comprise a heavy chaincomplementarity determining region 1 (CDR1) having the amino acid sequence of “SerSer Ser Tyr Tyr Trp Gly” (SEQ ID NO: 81); a heavy chain complementarity determiningregion 2 (CDR2) having the amino acid sequence of “Gly He Tyr Tyr Ser Gly Ser Thr TyrTyr Asn Pro Ser Leu Lys Ser” (SEQ ID NO: 82); and a heavy chain complementaritydetermining region 3 (CDR3) having the amino acid sequence of “Gin Arg Gly His SerSer Gly Trp Trp Tyr Phe Asp Leu” (SEQ ID NO: 83).

[0045] Further embodiments include fully human monoclonal antibodies, orbinding fragment thereof, having a light chain complementarity determining region 1(CDR1) having the amino acid sequence of “Arg Ala Ser Gin Gly He Ser Ser Tyr LeuAla” (SEQ ID NO: 84); a light chain complementarity determining region 2 (CDR2) 10 WO 2007/070432 PCT/US2006/047059 having the amino acid sequence of “Ala Ala Ser Ser Leu Gin Ser” (SEQ ID NO: 85); anda light chain complementarity determining region 3 (CDR3) having the amino acidsequence of “Gin Gin Ala Asn Asn Phe Pro Phe Thr” (SEQ ID NO: 86).

[0046] In other embodiments, the invention provides fully human monoclonalantibodies, or binding fragment thereof, that bind to insulin-like growth factor-II (IGF-II)with cross-reactivity to insulin-like growth factor-I (IGF-I), and comprise a heavy chaincomplementarity determining region 1 (CDR1) having the amino acid sequence of “SerSer Ser Asn Tyr Trp Gly” (SEQ ID NO: 87); a heavy chain complementarity determiningregion 2 (CDR2) having the amino acid sequence of “Gly lie Tyr Tyr Ser Gly Ser Thr TyrTyr Asn Pro Ser Leu Arg Ser” (SEQ ID NO: 88); and a heavy chain complementaritydetermining region 3 (CDR3) having the amino acid sequence of “Gin Arg Gly His SerSer Gly Trp Trp Tyr Phe Asp Leu” (SEQ ID NO: 89).

[0047] Further embodiments include fully human monoclonal antibodies, orbinding fragment thereof, having a light chain complementarity determining region 1(CDR1) having the amino acid sequence of “Arg Ala Ser Arg Gly lie Ser Ser Trp LeuAla” (SEQ ID NO: 90); a light chain complementarity determining region 2 (CDR2)having the amino acid sequence of “Thr Ala Ser Ser Leu Gin Ser” (SEQ ID NO: 91); anda light chain complementarity determining region 3 (CDR3) having the amino acidsequence of “Gin Gin Ala Asn Ser Phe Pro Phe Thr” (SEQ ID NO: 92).

[0048] Some embodiments provide the use of the specific binding proteinsdescribed herein in the preparation of a medicament for the treatment of a malignanttumor. In some aspects, the specific binding protein can be a fully human monoclonalantibody. In certain aspects, the binding protein is mAb 7.251.3 (ATCC AccessionNumber PTA-7422) or mAb 7.34.1 (ATCC Accession Number PTA-7423) or mAb7.159.2 (ATCC Accession Number PTA-7424). In some aspects, the medicament is foruse in combination with a second anti-neoplastic agent selected from the group consistingof an antibody, a chemotherapeutic agent, and a radioactive drug. In some aspects, themedicament is for use in conjunction with or following a conventional surgery, a bonemarrow stem cell transplantation or a peripheral stem cell transplantation.

[0049] The malignant tumor can be melanoma, non-small cell lung cancer,glioma, hepatocellular (liver) carcinoma, thyroid tumor, gastric (stomach) cancer,prostrate cancer, breast cancer, ovarian cancer, bladder cancer, lung cancer, glioblastoma,endometrial cancer, kidney cancer, colon cancer, pancreatic cancer, and epidermoidcarcinoma, for example. 11 WO 2007/070432 PCT/US2006/047059 [0050] Other embodiments provide the use of the specific binding proteinsdescribed herein in the preparation of a medicament for the treatment of a growth factor-dependent disease. In some aspects, the specific binding protein is a fully humanmonoclonal antibody and can be selected from the group consisting of mAh 7.251.3(ATCC Accession Number PTA-7422), mAb 7.34.1 (ATCC Accession Number PTA-7423), and mAb 7.159.2 (ATCC Accession Number PTA-7424).

[0051] The growth factor-dependent disease can be osteoporosis, diabetes, andcardiovascular diseases, for example.

[0052] Preferably, the antibody comprises a heavy chain amino acid sequencehaving a complementarity determining region (CDR) with one or more of the sequencesshown in Table 11. For example, the antibody can comprise a heavy chain amino acidsequence having the CDR1, CDR2, or CDR3 of one or more of the sequences shown inTable 11, or a combination thereof. It is noted that those of ordinary skill in the art canreadily accomplish CDR determinations. See for example, Kabat et al., Sequences ofProteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, BethesdaMD (1991), vols. 1-3.

[0053] Embodiments of the invention described herein relate to monoclonalantibodies that bind IGF-I/II and affect IGF-I/II function. Other embodiments relate tofully human anti-IGF-I/II antibodies and anti-IGF-I/II antibody preparations withdesirable properties from a therapeutic perspective, including high binding affinity forIGF-I/II, the ability to neutralize IGF-I/II in vitro and in vivo, and the ability to inhibitIGF-I/II induced cell proliferation.

BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 is a graph showing inhibition of xenograft tumor growth innude mice of NIH3T3 cells expressing IGF-II and IGF-IR (Clone 32 cells) with mAbs7.159.2, 7.34.1, 7.251.3 compared to IgG2 and PBS controls. Mean tumor volume isshown on the y-axis and time after implantation is shown on the x-axis.

[0055] Figure 2 is a graph showing body weight in Clone 32 xenograft micetreated with mAbs 7.159.2, 7.34.1, 7.251.3 compared to IgG2 and PBS controls. Meanbody weight is shown on the y-axis and time after implantation is shown on the x-axis.

[0056] Figure 3 is a graph showing inhibition of xenograft tumor growth innude mice of NIH3T3 cells expressing IGF-I and IGF-IR (P12 cells) with mAb 7.159.2 12 WO 2007/070432 PCT/US2006/047059 compared to PBS control. Mean tumor volume is shown on the y-axis and time after implantation (indicated by date) is shown on the x-axis.

DETAILED DESCRIPTION

[0057] Embodiments of the invention described herein relate to bindingproteins that specifically bind, to IGF-II with cross reactivity to IGF-I (referred to hereinas "IGFI/II"). In some embodiments, the binding proteins are antibodies, or bindingfragments thereof, and bind to IGF-II with cross-reactivity to IGF-I and inhibit thebinding of these proteins to their receptor, IGF-IR. Other embodiments of the inventioninclude fully human neutralizing anti-IGF-I/II antibodies, and antibody preparations thatare therapeutically useful and bind both insulin-like growth factors. Such anti-IGF-I/IIantibody preparations preferably have desirable therapeutic properties, including strongbinding affinity for IGF-I/II, the ability to neutralize IGF-I/II in vitro, and the ability toinhibit IGF-I/II-induced cell proliferation in vivo.

[0058] Embodiments of the invention also include isolated binding fragmentsof anti-IGF-I/II antibodies. Preferably, the binding fragments are derived from fullyhuman anti-IGF-I/II antibodies. Exemplary fragments include Fv, Fab’ or other wellknow antibody fragments, as described in more detail below. Embodiments of theinvention also include cells that express fully human antibodies against IGF-I/II.Examples of cells include hybridomas, or recombinantly created cells, such as Chinesehamster ovary (CHO) cells that produce antibodies against IGF-I/II.

[0059] In addition, embodiments of the invention include methods of usingthese antibodies for treating diseases. Anti-IGF-I/II antibodies are useful for preventingIGF-I/II mediated IGF-I/II signal transduction, thereby inhibiting cell proliferation. Themechanism of action of this inhibition may include inhibition of IGF-I/II from binding toits receptor, IGF-IR, inhibition of IGF-I/II induced IGF-IR signaling, or enhancedclearance of IGF-I/II therein lowering the effective concentration of IGF-I/II for bindingto IGF-IR. Diseases that are treatable through this inhibition mechanism include, but arenot limited to, neoplastic diseases, such as, melanoma, non-small cell lung cancer,glioma, hepatocellular (liver) carcinoma, gynecologic tumors, head and neck cancer,esophageal cancer, glioblastoma, and cancers and tumors of the thyroid, stomach,prostrate, breast, ovary, bladder, lung, uterus, kidney, colon, and pancreas, salivary gland,and colorectum. 13 WO 2007/070432 PCT/US2006/047059 [0060] Other embodiments of the invention include diagnostic assays forspecifically determining the quantity of IGF-I/II in a biological sample. The assay kit caninclude anti-IGF-Ι/Π antibodies along with the necessary labels for detecting suchantibodies. These diagnostic assays are useful to screen for growth factor-related diseasesincluding, but not limited to, neoplastic diseases, such as, melanoma, non-small cell lungcancer, glioma, hepatocellular (liver) carcinoma, gynecologic tumors, head and neckcancer, esophageal cancer, glioblastoma, and carcinoma of the thyroid, stomach,prostrate, breast, ovary, bladder, lung, uterus, kidney, colon, and pancreas, salivary gland,and. colorectum. Other non-neoplastic disease conditions may include acromegaly andgigantism, psoriasis, osteoporosis, atherosclerosis and smooth muscle restenosis of bloodvessels, as well as diabetes.

[0061] Further embodiments, features, and the like regarding anti-IGF-I/IIantibodies are provided in additional detail below.

Sequence Listing [0062] Embodiments of the invention include the specific anti-IGF-I/IIantibodies listed below in Table 1. This table reports the identification number of eachanti-IGF-Ι/Π antibody, along with the SEQ ID number of the corresponding heavy chainand light chain genes. Further, the germline sequences from which each heavy chain andlight chain derive are also provided below in Table 1.

[0063] Each antibody has been given an identification number that includeseither two or three numbers separated by one or two decimal points. In some cases,several clones of one antibody were prepared. Although the clones have the identicalnucleic acid and amino acid sequences as the parent sequence, they may also be listedseparately, with the clone number indicated by the number to the right of a seconddecimal point. Thus, for example, the nucleic acid and amino acid sequences of antibody7.159.2 are identical to the sequences of antibody 7.159.1.

[0064] As can be seen by comparing the sequences in the sequence listing,SEQ ID NOs.: 1-20 differ from SEQ ID NOs.: 39-58 because SEQ ID NOs.: 39-58include the untranslated, signal peptide, and constant domain regions for each sequencedheavy or light chain. 14 WO 2007/070432 PCT/US2006/047059 TABLE 1 mAb IDNo.: Sequence SEQ ID NO: 7.158.1 Nucleotide sequence encoding the variable region of the heavy chain 1 Amino acid sequence encoding the variable region of the heavy chain 2 Nucleotide sequence encoding the variable region of the light chain 3 Amino acid sequence encoding the variable region of the light chain 4 7.159.2 Nucleotide sequence encoding the variable region of the heavy chain 5 Amino acid sequence encoding the variable region of the heavy chain 6 Nucleotide sequence encoding the variable region of the light chain 7 Amino acid sequence encoding the variable region of the light chain 8 7.34.1 Nucleotide sequence encoding the variable region of the heavy chain 9 Amino acid sequence encoding the variable region of the heavy chain 10 Nucleotide sequence encoding the variable region of the light chain 11 Amino acid sequence encoding the variable region of the light chain 12 7.251.3 Nucleotide sequence encoding the variable region of the heavy chain 13 Amino acid sequence encoding the variable region of the heavy chain 14 Nucleotide sequence encoding the variable region of the light chain 15 Amino acid sequence encoding the variable region of the light chain 16 7.234.1 Nucleotide sequence encoding the variable region of the heavy chain 17 Amino acid sequence encoding the variable region of the heavy chain 18 Nucleotide sequence encoding the variable region of the light chain 19 Amino acid sequence encoding the variable region of the light chain 20 7.158.1 Nucleotide sequence encoding the variable region of the heavy chain 39 Amino acid sequence encoding the variable region of the heavy chain 40 Nucleotide sequence encoding the variable region of the light chain 41 Amino acid sequence encoding the variable region of the light chain 42 7.159.2 Nucleotide sequence encoding the variable region of the heavy chain 43 Amino acid sequence encoding the variable region of the heavy chain 44 Nucleotide sequence encoding the variable region of the light chain 45 Amino acid sequence encoding the variable region of the light chain 46 7.34.1 Nucleotide sequence encoding the variable region of the heavy chain 47 Amino acid sequence encoding the variable region of the heavy chain 48 Nucleotide sequence encoding the variable region of the light chain 49 Amino acid sequence encoding the variable region of the light chain 50 7.251.3 Nucleotide sequence encoding the variable region of the heavy chain 51 Amino acid sequence encoding the variable region of the heavy chain 52 Nucleotide sequence encoding the variable region of the light chain 53 Amino acid sequence encoding the variable region of the light chain 54 7.234.1 Nucleotide sequence encoding the variable region of the heavy chain 55 Amino acid sequence encoding the variable region of the heavy chain 56 Nucleotide sequence encoding the variable region of the light chain 57 Amino acid sequence encoding the variable region of the light chain 58 Germline (7.158.1) Nucleotide sequence encoding the variable region of the heavy chain 59 Amino acid sequence encoding the variable region of the heavy chain 60 Nucleotide sequence encoding the variable region of the light chain 61 15 WO 2007/070432 PCT/US2006/047059

Amino acid sequence encoding the variable region of the light chain 62 Germline (7.159.1) Nucleotide sequence encoding the variable region of the heavy chain 63 Amino acid sequence encoding the variable region of the heavy chain 64 Nucleotide sequence encoding the variable region of the light chain 65 Amino acid sequence encoding the variable region of the light chain 66 Germline (7.34.1) Nucleotide sequence encoding the variable region of the heavy chain 67 Amino acid sequence encoding the variable region of the heavy chain 68 Nucleotide sequence encoding the variable region of the light chain 69 Amino acid sequence encoding the variable region of the light chain 70 Germline (7.251.3) Nucleotide sequence encoding the variable region of the heavy chain 71 Amino acid sequence encoding the variable region of the heavy chain 72 Nucleotide sequence encoding the variable region of the light chain 73 Amino acid sequence encoding the variable region of the light chain 74

Definitions [0065] Unless otherwise defined, scientific and technical terms used hereinshall have the meanings that are commonly understood by those of ordinary skill in theart. Further, unless otherwise required by context, singular terms shall include pluralitiesand plural terms shall include the singular. Generally, nomenclatures utilized inconnection with, and techniques of, cell and tissue culture, molecular biology, and proteinand oligo- or polynucleotide chemistry and hybridization described herein are those wellknown and commonly used in the art.

[0066] Standard techniques are used for recombinant DNA, oligonucleotidesynthesis, and tissue culture and transformation (e.g., electroporation, lipofection).Enzymatic reactions and purification techniques are performed according tomanufacturer’s specifications or as commonly accomplished in the art or as describedherein. The foregoing techniques and procedures are generally perfonned according toconventional methods well known in the art and as described in various general and morespecific references that are cited and discussed throughout the present specification. Seee.g., Sambrook et al. Molecular Cloning: A Laboratory Manual (3rd ed., Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y. (2001)), which is incorporated hereinby reference. The nomenclatures utilized in connection with, and the laboratoryprocedures and techniques of, analytical chemistry, synthetic organic chemistry, andmedicinal and pharmaceutical chemistry described herein are those well known andcommonly used in the art. Standard techniques are used for chemical syntheses, chemicalanalyses, pharmaceutical preparation, formulation, and delivery, and treatment ofpatients. 16 WO 2007/070432 PCT/US2006/047059 [0067] As utilized in accordance with the present disclosure, the followingterms, unless otherwise indicated, shall be understood to have the following meanings: [0068] The term “IGF-I” refers to the molecule Insulin-like growth factor-I,and the term “IGF-Π” refers to the molecule Insulin-like growth factor-II. The term“IGF-I/II” refers to both molecules Insulin-like growth factors-I and -II, and relates to thepreferential binding to IGF-II with cross-reactivity to IGF-I. Thus, an antibody that bindsto IGF-I/II will preferentially bind to IGF-II, but would cross-react with IGF-I, binding toIGF-II with higher affinity than to IGF-I. For example, the antibody can bind to IGF-IIwith 2.5 times greater affinity than to IGF-I. In certain embodiments, the antibody canbind to IGF-II with at least 5, at least 10, at least 25, at least 50 or at least 150 timesgreater affinity than to IGF-I.

[0069] The term “neutralizing” when referring to an antibody relates to theability of an antibody to eliminate, or significantly reduce, the activity of a target antigen.Accordingly, a “neutralizing” anti-IGF-I/II antibody is capable of eliminating orsignificantly reducing the activity of IGF-I/II. A neutralizing IGF-I/II antibody may, forexample, act by blocking the binding of IGF-I/II to its receptor IGF-IR. By blocking thisbinding, the IGF-IR mediated signal transduction is significantly, or completely,eliminated. Ideally, a neutralizing antibody against IGF-I/II inhibits cell proliferation.

[0070] The term “isolated polynucleotide” as used herein shall mean apolynucleotide that has been isolated from its naturally occurring environment. Suchpolynucleotides may be genomic, cDNA, or synthetic. Isolated polynucleotidespreferably are not associated with all or a portion of the polynucleotides they associatewith in nature. The isolated polynucleotides may be operably linked to anotherpolynucleotide that it is not linked to in nature. In addition, isolated polynucleotidespreferably do not occur in nature as part of a larger sequence.

[0071] The term “isolated protein” referred to herein means a protein that hasbeen isolated from its naturally occurring environment. Such proteins may be derivedfrom genomic DNA, cDNA, recombinant DNA, recombinant RNA, or synthetic origin orsome combination thereof, which by virtue of its origin, or source of derivation, the“isolated protein” (1) is not associated with proteins found in nature, (2) is free of otherproteins from the same source, e.g. free of murine proteins, (3) is expressed by a cell froma different species, or (4) does not occur in nature.

[0072] The term “polypeptide” is used herein as a generic term to refer tonative protein, fragments, or analogs of a polypeptide sequence. Hence, native protein, 17 WO 2007/070432 PCT/US2006/047059 fragments, and analogs are species of the polypeptide genus. Preferred polypeptides inaccordance with the invention comprise the human heavy chain immunoglobulinmolecules and the human kappa light chain immunoglobulin molecules, as well asantibody molecules formed by combinations comprising the heavy chain immunoglobulinmolecules with light chain immunoglobulin molecules, such as the kappa or lambda lightchain immunoglobulin molecules, and vice versa, as well as fragments and analogsthereof. Preferred polypeptides in accordance with the invention may also comprisesolely the human heavy chain immunoglobulin molecules or fragments thereof.

[0073] The term “naturally-occurring” as used herein as applied to an objectrefers to the fact that an object can be found in nature. For example, a polypeptide orpolynucleotide sequence that is present in an organism (including viruses) that can beisolated from a source in nature and which has not been intentionally modified by man inthe laboratory or otherwise is naturally-occurring.

[0074] The term “operably linked” as used herein refers to positions ofcomponents so described that are in a relationship permitting them to function in theirintended manner. For example, a control sequence “operably linked” to a codingsequence is connected in such a way that expression of the coding sequence is achievedunder conditions compatible with the control sequences.

[0075] The term “polynucleotide” as referred to herein means a polymericform of nucleotides of at least 10 bases in length, either ribonucleotides ordeoxynucleotides or a modified form of either type of nucleotide, or RNA-DNA hetero-duplexes. The term includes single and double stranded forms of DNA.

[0076] The term “oligonucleotide” referred to herein includes naturallyoccurring, and modified nucleotides linked together by naturally occurring, and non-naturally occurring linkages. Oligonucleotides are a polynucleotide subset generallycomprising a length of 200 bases or fewer. Preferably, oligonucleotides are 10 to 60bases in length and most preferably 12, 13, 14, 15, 16, 17, 18, 19, or 20 to 40 bases inlength. Oligonucleotides are usually single stranded, e.g. for probes; althougholigonucleotides may be double stranded, e.g. for use in the construction of a genemutant. Oligonucleotides can be either sense or antisense oligonucleotides.

[0077] The term “naturally occurring nucleotides” referred to herein includesdeoxyribonucleotides and ribonucleotides. The term “modified nucleotides” referred toherein includes nucleotides with modified or substituted sugar groups and the like. Theterm “oligonucleotide linkages” referred to herein includes oligonucleotides linkages such 18 WO 2007/070432 PCT/US2006/047059 as phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate,phosphoroanilothioate, phosphoraniladate, phosphoroamidate, and the like. See e.g.,LaPlanche et al. Nucl. Acids Res. 14:9081 (1986); Stec et al. J. Am. Chem. Soc. 106:6077(1984); Stein et al. Nucl. Acids Res. 16:3209 (1988); Zon et al. Anti-Cancer Drug Design6:539 (1991); Zon et al. Oligonucleotides and Analogies: A Practical Approach, pp. 87-108 (F. Eckstein, Ed., Oxford University Press, Oxford England (1991)); Stec et al. U.S.Patent No. 5,151,510; Uhlmann and Peyman Chemical Reviews 90:543 (1990), thedisclosures of which are hereby incorporated by reference. An oligonucleotide caninclude a label for detection, if desired.

[0078] The term “selectively hybridize” referred to herein means to detectablyand specifically bind. Polynucleotides, oligonucleotides and fragments thereofselectively hybridize to nucleic acid strands under hybridization and wash conditions thatminimize appreciable amounts of detectable binding to nonspecific nucleic acids. Highstringency conditions can be used to achieve selective hybridization conditions as knownin the art and discussed herein. Generally, the nucleic acid sequence homology betweenthe polynucleotides, oligonucleotides, or antibody fragments and a nucleic acid sequenceof interest will be at least 80%, and more typically with preferably increasing homologiesof at least 85%, 90%, 95%, 99%, and 100%. ]0079] Two amino acid sequences are “homologous” if there is a partial orcomplete identity between their sequences. For example, 85% homology means that 85%of the amino acids are identical when the two sequences are aligned for maximummatching. Gaps (in either of the two sequences being matched) are allowed inmaximizing matching; gap lengths of 5 or less are preferred with 2 or less being morepreferred. Alternatively and preferably, two protein sequences (or polypeptide sequencesderived from them of at least about 30 amino acids in length) are homologous, as thisterm is used herein, if they have an alignment score of at more than 5 (in standarddeviation units) using the program ALIGN with the mutation data matrix and a gappenalty of 6 or greater. See Dayhoff, M.O., in Atlas of Protein Sequence and Structure,pp. 101-110 (Volume 5, National Biomedical Research Foundation (1972)) andSupplement 2 to this volume, pp. 1-10. The two sequences or parts thereof are morepreferably homologous if their amino acids are greater than or equal to 50% identicalwhen optimally aligned using the ALIGN program. It should be appreciated that therecan be differing regions of homology within two orthologous sequences. For example, 19 WO 2007/070432 PCT/US2006/047059 the functional sites of mouse and human orthologues may have a higher degree ofhomology than non-functional regions.

[0080] The term “corresponds to” is used herein to mean that a polynucleotidesequence is homologous (i.e., is identical, not strictly evolutionarily related) to all or aportion of a reference polynucleotide sequence, or that a polypeptide sequence is identicalto a reference polypeptide sequence.

[0081] In contradistinction, the term “complementary to” is used herein tomean that the complementary sequence is homologous to all or a portion of a referencepolynucleotide sequence. For illustration, the nucleotide sequence “TATAC”corresponds to a reference sequence “TATAC” and is complementary to a referencesequence “GTATA”.

[0082] The following terms are used to describe the sequence relationshipsbetween two or more polynucleotide or amino acid sequences: “reference sequence”,“comparison window”, “sequence identity”, “percentage of sequence identity”, and“substantial identity”. A “reference sequence” is a defined sequence used as a basis for asequence comparison. A reference sequence may be a subset of a larger sequence, forexample, as a segment of a full-length cDNA or gene sequence given in a sequence listingor may comprise a complete cDNA of gene sequence. Generally, a reference sequence isat least 18 nucleotides or 6 amino acids in length, frequently at least 24 nucleotides or 8amino acids in length, and often at least 48 nucleotides or 16 amino acids in length. Sincetwo polynucleotides or amino acid sequences may each (1) comprise a sequence (i.e., aportion of the complete polynucleotide or amino acid sequence) that is similar betweenthe two molecules, and (2) may further comprise a sequence that is divergent between thetwo polynucleotides or amino acid sequences, sequence comparisons between two (ormore) molecules are typically performed by comparing sequences of the two moleculesover a “comparison window” to identify and compare local regions of sequencesimilarity. A “comparison window”, as used herein, refers to a conceptual segment of atleast about 18 contiguous nucleotide positions or about 6 amino acids wherein thepolynucleotide sequence or amino acid sequence is compared to a reference sequence ofat least 18 contiguous nucleotides or 6 amino acid sequences and wherein the portion ofthe polynucleotide sequence in the comparison window may include additions, deletions,substitutions, and the like (i.e., gaps) of 20 percent or less as compared to the referencesequence (which does not comprise additions or deletions) for optimal alignment of thetwo sequences. Optimal alignment of sequences for aligning a comparison window may 20 191718/2 be conducted by the local homology algorithm of Smith and Waterman Adv. Appl. Math.2:482 (1981), by the homology alignment algorithm of Needleman and Wunsch J. Mol.Biol. 48:443 (1970), by the search for similarity method of Pearson and Lipman Proc. NatlAcad. Sci. (U.S.A.) 85:2444 (1988), by computerized implementations of these algorithms(GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software PackageRelease 7.0, (Genetics Computer Group, 575 Science Dr., Madison, Wis.),GENEWORKS™, or MACVECTOR® software packages), or by inspection, and the bestalignment (i.e., resulting in the highest percentage of homology over the comparisonwindow) generated by the various methods is selected.

[0083] The term "sequence identity" means that two polynucleotide or aminoacid sequences are identical (i.e., on a nucleotide-by-nucleotide or residue-by-residuebasis) over the comparison window. The term "percentage of sequence identity" iscalculated by comparing two optimally aligned sequences over the window of comparison,determining the number of positions at which the identical nucleic acid base (e.g., A, T, C,G, U, or I) or amino acid residue occurs in both sequences to yield the number of matchedpositions, dividing the number of matched positihns by the total number of positions in thecomparison window (i.e., the window size), and multiplying the result by 100 to yield thepercentage of sequence identity. The terms "substantial identity" as used herein denotes acharacteristic of a polynucleotide or amino acid sequence, wherein the polynucleotide oramino acid comprises a sequence that has at least 85 percent sequence identity, preferablyat least 90 to 95 percent sequence identity, more preferably at least 99 percent sequenceidentity, as compared to a reference sequence over a comparison window of at least 18nucleotide (6 amino acid) positions, frequently over a window of at least 24-48 nucleotide(8-16 amino acid) positions, wherein the percentage of sequence identity is calculated bycomparing the reference sequence to the sequence which may include deletions oradditions which total 20 percent or less of the reference sequence over the comparisonwindow. The reference sequence may be a subset of a larger sequence.

[0084] As used herein, the twenty conventional amino acids andtheir abbreviations follow conventional usage. See Immunology - A Synthesis (2nd Edition,E.S. Golub and D.R. Gren, Eds., Shiauer Associates, Sunderland, Mass.(1991)). Stereoisomers (e.g., D-amino acids) of the twenty conventionalamino acids, unnatural amino acids such as α-, α-disubstituted amino acids, 21 P/l 5804/125647/596826/1 WO 2007/070432 PCT/US2006/047059 N-alkyl amino acids, lactic acid, and other unconventional amino acids may also besuitable components for polypeptides of the present invention. Examples ofunconventional amino acids include: 4-hydroxyproline, γ-carboxyglutamate, ε-Ν,Ν,Ν-trimethyllysine, ε-Ν-acetyllysine, O-phosphoserine, N-acetylserine, N-formylmethionine,3-methylhistidine, 5-hydroxylysine, σ-Ν-methylarginine, and other similar amino acidsand imino acids (e.g., 4-hydroxyproline). In the polypeptide notation used herein, theleft-hand direction is the amino terminal direction and the right-hand direction is thecarboxy-terminal direction, in accordance with standard usage and convention.

[0085] Similarly, unless specified otherwise, the left-hand end of single-stranded polynucleotide sequences is the 5’ end; the left-hand direction of double-stranded polynucleotide sequences is referred to as the 5’ direction. The direction of 5’ to3’ addition of nascent RNA transcripts is referred to as the transcription direction;sequence regions on the DNA strand having the same sequence as the RNA and whichare 5’ to the 5’ end of the RNA transcript are referred to as “upstream sequences”;sequence regions on the DNA strand having the same sequence as the RNA and whichare 3’ to the 3’ end of the RNA transcript are referred to as “downstream sequences”.

[0086] As applied to polypeptides, the term “substantial identity” means thattwo peptide sequences, when optimally aligned, such as by the programs GAP orBESTFIT using default gap weights, share at least 80 percent sequence identity,preferably at least 90 percent sequence identity, more preferably at least 95 percentsequence identity, and most preferably at least 99 percent sequence identity. Preferably,residue positions that are not identical differ by conservative amino acid substitutions.Conservative amino acid substitutions refer to the interchangeability of residues havingsimilar side chains. For example, a group of amino acids having aliphatic side chains isglycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic-hydroxyl side chains is serine and threonine; a group of amino acids having amide-containing side chains is asparagine and glutamine; a group of amino acids havingaromatic side chains is phenylalanine, tyrosine, and tryptophan; a group of amino acidshaving basic side chains is lysine, arginine, and histidine; and a group of amino acidshaving sulfur-containing side chains is cysteine and methionine. Preferred conservativeamino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine,lysine-arginine, alanine-valine, glutamic-aspartic, and asparagine-glutamine. 22 WO 2007/070432 PCT/US2006/047059 [0087] As discussed herein, minor variations in the amino acid sequences ofantibodies or immunoglobulin molecules are contemplated as being encompassed by thepresent invention, providing that the variations in the amino acid sequence maintain atleast 75%, more preferably at least 80%, 90%, 95%, and most preferably 99% sequenceidentity to the antibodies or immunoglobulin molecules described herein. In particular,conservative amino acid replacements are contemplated. Conservative replacements arethose that take place within a family of amino acids that have related side chains.Genetically encoded amino acids are generally divided into families: (1) acidic=aspartate,glutamate; (2) basic=lysine, arginine, histidine; (3) non-polar=alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine, tryptophan; and (4) unchargedpolar=glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine. Morepreferred families are: serine and threonine are an aliphatic-hydroxy family; asparagineand glutamine are an amide-containing family; alanine, valine, leucine and isoleucine arean aliphatic family; and phenylalanine, tryptophan, and tyrosine are an aromatic family.For example, it is reasonable to expect that an isolated replacement of a leucine with anisoleucine or valine, an aspartate with a glutamate, a threonine with a serine, or a similarreplacement of an amino acid with a structurally related amino acid will not have a majoreffect on the binding function or properties of the resulting molecule, especially if thereplacement does not involve an amino acid within a framework site. Whether an aminoacid change results in a functional peptide can readily be determined by assaying thespecific activity of the polypeptide derivative. Assays are described in detail herein.Fragments or analogs of antibodies or immunoglobulin molecules can be readily preparedby those of ordinary skill in the art. Preferred amino- and carboxy-termini of fragmentsor analogs occur near boundaries of functional domains. Structural and functionaldomains can be identified by comparison of the nucleotide and/or amino acid sequencedata to public or proprietary sequence databases. Preferably, computerized comparisonmethods are used to identify sequence motifs or predicted protein conformation domainsthat occur in other proteins of known structure and/or function. Methods to identifyprotein sequences that fold into a known three-dimensional structure are known. Bowieet al. Science 253:164 (1991). Thus, the foregoing examples demonstrate that those ofskill in the art can recognize sequence motifs and structural conformations that may beused to define structural and functional domains in accordance with the antibodiesdescribed herein. 23 191718/2 [0088] Preferred amino acid substitutions are those which: (1) reducesusceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinityfor forming protein complexes, (4) alter binding affinities, and (5) confer or modify otherphysicochemical or functional properties of such analogs. Analogs can include variousmuteins of a sequence other than the naturally-occurring peptide sequence. For example,single or multiple amino acid substitutions (preferably conservative amino acidsubstitutions) may be made in the naturally-occurring sequence (preferably in the portionof the polypeptide outside the domain(s) forming intermolecular contacts. A conservativeamino acid substitution should not substantially change the structural characteristics of theparent sequence (e.g., a replacement amino acid should not tend to break a helix thatoccurs in the parent sequence, or disrupt other types of secondary structure thatcharacterizes the parent sequence). Examples of art-recognized polypeptide secondary andtertiary structures are described in Proteins, Structures and Molecular Principles(Creighton, Ed., W. H. Freeman and Company, New York (1984)); Introduction to ProteinStructure (C. Branden and J. Tooze, eds., Garland Publishing, New York, N. Y. (1991));and Thornton et al Nature 354:105 (1991).

[0089] The term "polypeptide fragment" as used herein refers to apolypeptide that has an amino-terminal and/or carboxy-terminal deletion, but where theremaining amino acid sequence is identical to the corresponding positions in the naturally-occurring sequence deduced, for example, from a full-length cDNA sequence. Fragmentstypically are at least 5, 6, 8 or 10 amino acids long, preferably at least 14 amino acids long,more preferably at least 20 amino acids long, usually at least 50 amino acids long, andeven more preferably at least 70 amino acids long. The term "analog" as used herein refersto polypeptides which are comprised of a segment of at least 25 amino acids that hassubstantial identity to a portion of a deduced amino acid sequence and which has at leastone of the following properties: (1) specific binding to IGF-I/II, under suitable bindingconditions, (2) ability to block appropriate IGF-I/II binding, or (3) ability to inhibit IGF-I/II activity. Typically, polypeptide analogs comprise a conservative amino acidsubstitution (or addition or deletion) with respect to the naturally-occurring sequence.Analogs typically are at least 20 amino acids long, preferably at least 50 amino acids longor longer, and can often be as long as a full-length naturally-occurring polypeptide [0090] Peptide analogs are commonly used in the pharmaceutical industry asnon-peptide drugs with properties analogous to those of the template peptide. These 24 P/l 5804/125647/596826/1 191718/2 types of non-peptide compound are termed "peptide mimetics" or "peptidomimetics".Fauchere, J. Adv. Drug Res. 15:29 (1986); Veber and Freidinger TINS p.392 (1985); andEvans et al. J. Med. Chem. 30:1229 (1987). Such compounds are often developed with theaid of computerized molecular modeling. Peptide mimetics that are structurally similar totherapeutically useful peptides may be used to produce an equivalent therapeutic orprophylactic effect. Generally, peptidomimetics are structurally similar to a paradigmpolypeptide (i.e., a polypeptide that has a biochemical property or pharmacologicalactivity), such as human antibody, but have one or more peptide linkages optionallyreplaced by a linkage selected from the group consisting of: -CH2NH~, ~CH2S~, -CH2-CH2~, -CH=CH-(cis and trans), - COCH2~, ~CH(OH)CH2~, and ~CH2SO~, bymethods well known in the art. Systematic substitution of one or more amino acids of aconsensus sequence with a D-amino acid of the same type (e.g., D-lysine in place of L-lysine) may be used to generate more stable peptides. In addition, constrained peptidescomprising a consensus sequence or a substantially identical consensus sequence variationmay be generated by methods known in the art (Rizo and Gierasch Ann. Rev. Biochem.61:387 (1992)); for example, by adding internal cysteine residues capable of formingintramolecular disulfide bridges which cyclize the peptide.

[0091] As used herein, the term "antibody" refers to a polypeptide or group ofpolypeptides that are comprised of at least one binding domain that is formed from thefolding of polypeptide chains having three-dimensional binding spaces with internalsurface shapes and charge distributions complementary to the features of an antigenicdeterminant of an antigen. An antibody typically has a tetrameric form, comprising twoidentical pairs of polypeptide chains, each pair having one "light" and one "heavy" chain.The variable regions of each light/heavy chain pair form an antibody binding site.

[0092] "Binding fragments" of an antibody are produced by recombinant DNAtechniques, or by enzymatic or chemical cleavage of intact antibodies. Binding fragmentsinclude Fab, Fab', F(ab')2, Fv, and single-chain antibodies. An antibody other than a"bispecific" or "bifunctional" antibody is understood to have each of its binding sitesidentical. An antibody substantially inhibits adhesion of a receptor to a counterreceptorwhen an excess of antibody reduces the quantity of receptor bound to counterreceptor by atleast about 20%, 40%, 60% or 80%, and more usually greater than about 85% (asmeasured in an in vitro competitive binding assay). 25 P/l 5804/125647/596826/1 WO 2007/070432 PCT/US2006/047059 [0093] As used herein, a “binding protein” or a “specific binding protein” areproteins that specifically bind to a target molecule. Antibodies, and binding fragments ofantibodies, are binding proteins.

[0094] The term “epitope” includes any protein determinant capable ofspecific binding to an immunoglobulin or T-cell receptor. Epitopic determinants usuallyconsist of chemically active surface groupings of molecules such as amino acids or sugarside chains and may, but not always, have specific three-dimensional structuralcharacteristics, as well as specific charge characteristics. An antibody is said tospecifically bind an antigen when the dissociation constant is <1 μΜ, preferably <100nM and most preferably <10 nM.

[0095] The term “agent” is used herein to denote a chemical compound, amixture of chemical compounds, a biological macromolecule, or an extract made frombiological materials.

[0096] “Active” or “activity” in regard to an IGF-I/II polypeptide refers to aportion of an IGF-I/II polypeptide that has a biological or an immunological activity of anative IGF-I/II polypeptide. “Biological” when used herein refers to a biological functionthat results from the activity of the native IGF-I/II polypeptide. A preferred IGF-I/IIbiological activity includes, for example, IGF-I/II induced cell proliferation. 10097] “Mammal” when used herein refers to any animal that is considered amammal. Preferably, the mammal is human.

[0098] Digestion of antibodies with the enzyme, papain, results in twoidentical antigen-binding fragments, known also as “Fab” fragments, and a “Fc”fragment, having no antigen-binding activity but having the ability to crystallize.Digestion of antibodies with the enzyme, pepsin, results in the a F(ab’)2 fragment inwhich the two arms of the antibody molecule remain linked and comprise two-antigenbinding sites. The F(ab’)2 fragment has the ability to crosslink antigen.

[0099] “Fv” when used herein refers to the minimum fragment of an antibodythat retains both antigen-recognition and antigen-binding sites.

[0100] “Fab” when used herein refers to a fragment of an antibody thatcomprises the constant domain of the light chain and the CHI domain of the heavy chain.

[0101] The term “mAb” refers to monoclonal antibody. 26 191718/2 [0102] "Liposome" when Used herein refers to a small vesicle that may be usefulfor delivery of drugs that may include the IGF-I/II polypeptide of the invention orantibodies to such an IGF-I/II polypeptide to a mammal.

[0103] "Label" or "labeled" as used herein refers to the addition of a detectablemoiety to a polypeptide, for example, a radiolabel, fluorescent label, enzymatic labelchemiluminescent labeled or a biotinyl group. Radioisotopes or radionuclides may include3H, 14C, I5N 35S, 90Y, 99Tc, 1HIn, 125I, 131I, fluorescent labels may include rhodamine,lanthanide phosphors or FITC and enzymatic labels may include horseradish peroxidase,β-galactosidase, luciferase, alkaline phosphatase.

[0104] The term "pharmaceutical agent or drug" as used herein refers to achemical compound or composition capable of inducing a desired therapeutic effect whenproperly administered to a patient. Other chemistry terms herein are used according toconventional usage in the art, as exemplified by The McGraw-Hill Dictionary of ChemicalTerms (Parker, S., Ed., McGraw-Hill, San Francisco (1985)).

[0105] As used herein, "substantially pure" means an object species is thepredominant species present (i.e., on a molar basis it is more abundant than any otherindividual species in the composition), and preferably a substantially purified fraction is acomposition wherein the object species comprises at least about 50 percent (on a molarbasis) of all macromolecular species present. Generally, a substantially pure compositionwill comprise more than about 80 percent of all macromolecular species present in thecomposition, more preferably more than about 85%, 90%, 95%, and 99%. Most preferably,the object species is purified to essential homogeneity (contaminant species cannot bedetected in the composition by conventional detection methods) wherein the compositionconsists essentially of a single macromolecular species.

[0106] The term "patient" includes human and veterinary subjects.

Human Antibodies and Humanization of Antibodies [0107] Human antibodies avoid some of the problems associated with antibodiesthat possess murine or rat variable and/or constant regions. The presence of such murine orrat derived proteins can lead to the rapid clearance of the antibodies or can lead to thegeneration of an immune response against the antibody by a patient. In order to avoid theutilization of murine or rat derived antibodies, fully human antibodies can be generatedthrough the introduction of functional human antibody loci into a rodent, other 27 P/l 5804/125647/596826/1 191718/2 mammal or animal so that the rodent, other mammal or animal produces fully humanantibodies.

[0108] One method for generating fully human antibodies is through the use ofXenoMouse® strains of mice that have been engineered to contain up to but less than 1000kb-sized germline configured fragments of the human heavy chain locus and kappa lightchain locus. See Mendez et al. Nature Genetics 15:146-156 (1997) and Green andJakobovits J. Exp. Med. 188:483-495 (1998). The XenoMouse® strains are available fromAbgenix, Inc. (Fremont, CA).

[0109] The production of the XenoMouse® strains of mice is further discussedand delineated in U.S. Patent Application Serial Nos. 07/466,008, filed January 12, 1990,07/610,515, filed November 8, 1990, 07/919,297, filed July 24, 1992, 07/922,649, filedJuly 30, 1992, 08/031,801, filed March 15, 1993, 08/112,848, filed August 27, 1993,08/234,145, filed April 28, 1994, 08/376,279, filed January 20, 1995, 08/430, 938, filedApril 27, 1995, 08/464,584, filed June 5, 1995, 08/464,582, filed June 5, 1995, 08/463,191,filed June 5, 1995, 08/462,837, filed June 5, 1995, 08/486,853, filed June 5, 1995,08/486,857, filed June 5, 1995, 08/486,859, filed June 5, 1995, 08/462,513, filed June 5,1995, 08/724,752, filed October 2, 1996, 08/759,620, filed December 3, 1996, U.S.Publication 2003/0093820, filed November 30, 2001 and U.S. Patent Nos. 6,162,963,6,150,584, 6,1 14,598, 6,075,181, and 5,939,598 and Japanese Patent Nos. 3 068 180 B2, 3068 506 B2, and 3 068 507 B2. See also European Patent No., EP 0 463 151 BI, grantpublished June 12, 1996, International Patent Application No., WO 94/02602, publishedFebruary 3, 1994, International Patent Application No., WO 96/34096, published October31, 1996, WO 98/24893, published June 11, 1998, WO 00/76310, published December 21,2000.

[0110] In an alternative approach, others, including GenPharm International,Inc., have utilized a "minilocus" approach. In the minilocus approach, an exogenous Iglocus is mimicked through the inclusion of pieces (individual genes) from the Ig locus.Thus, one or more Vh genes, one or more Dh genes, one or more Jh genes, a mu constantregion, and usually a second constant region (preferably a gamma constant region) areformed into a construct for insertion into an animal. This approach is described in U.S.Patent No. 5,545,807 to Surani et al. and U.S. Patent Nos. 5,545,806, 5,625,825,5,625,126, 5,633,425, 5,661,016, 5,770,429, 5,789,650, 5,814,318, 5,877,397, 5,874,299, 28 P/15804/125647/596826/l 191718/2 and 6,255,458 each to Lonberg and Kay, U.S. Patent No. 5,591,669 and 6,023.010 toKrimpenfort and Bems, U.S. Patent Nos. 5,612,205, 5,721,367, and 5,789,215 to Bems etal, and U.S. Patent No. 5,643,763 to Choi and Dunn, and GenPharm International U.S.Patent Application Serial Nos. 07/574,748, filed August 29, 1990, 07/575,962, filedAugust 31, 1990, 07/810,279, filed December 17, 1991, 07/853,408, filed March 18,1992,07/904,068, filed June 23, 1992, 07/990,860, filed December 16, 1992, 08/053,131, filedApril 26, 1993, 08/096,762, filed July 22, 1993, 08/155,301, filed November 18, 1993,08/161 ,739, filed December 3, 1993, 08/165,699, filed December 10, 1993, 08/209,741,filed March 9, 1994. See also European Patent No. 0 546 073 BI, International PatentApplication Nos. WO 92/03918, WO 92/22645, WO 92/22647, WO 92/22670, WO93/12227, WO 94/00569, WO 94/25585, WO 96/14436, WO 97/13852, and WO 98/24884and U.S. Patent No. 5,981,175. See further Taylor et al., 1992, Chen et al., 1993, Tuaillonet al., 1993, Choi et al., 1993, Lonberg et al., (1994), Taylor et al., (1994), and Tuaillon etal., (1995), Fishwild et al., (1996).

[0111] Kirin has also demonstrated the generation of human antibodies frommice in which, through microcell fusion, large pieces of chromosomes, or entirechromosomes, have been introduced. See European Patent Application Nos. 773 288 and843 961. Additionally, KM™-mice, which are the result of cross-breeding of Kirin's Tcmice with Medarex's minilocus (Humab) mice have been generated. These mice possessthe human IgH transchromosome of the Kirin mice and the kappa chain transgene of theGenpharm mice (Ishida et al., Cloning Stem Cells, (2002) 4:91-102).

[0112] Human antibodies can also be derived by in vitro methods. Suitableexamples include but are not limited to phage display (CAT, Morphosys, Dyax,Biosite/Medarex, Xoma, Symphogen, Alexion (formerly Proliferon), Affimed) ribosomedisplay (CAT), yeast display, and the like.

Preparation of Antibodies [0113] Antibodies, as described herein, were prepared through the utilizationof the XenoMouse® technology, as described below. Such mice, then, are capable of 29 P/l 5804/125647/596826/1 191718/2 producing human immunoglobulin molecules and antibodies and are deficient in theproduction of murine immunoglobulin molecules and antibodies. Technologies utilized forachieving the same are disclosed in the patents, applications, and references disclosed inthe background section herein. In particular, however, a preferred embodiment oftransgenic production of mice and antibodies therefrom is disclosed in U.S. PatentApplication Serial No. 08/759,620, filed December 3, 1996 and International PatentApplication Nos. WO 98/24893, published June 11, 1998 and WO 00/76310, publishedDecember 21, 2000. See also Mendez etal. Nature Genetics 15:146-156 (1997).

[0114] Through the use of such technology, fully human monoclonal antibodiesto a variety of antigens have been produced. Essentially, XenoMouse® lines of mice areimmunized with an antigen of interest (e.g. IGF-I/II), lymphatic cells (such as B-cells) arerecovered from the hyper-immunized mice, and the recovered lymphocytes are fused witha myeloid-type cell line to prepare immortal hybridoma cell lines. These hybridoma celllines are screened and selected to identify hybridoma cell lines that produced antibodiesspecific to the antigen of interest. Provided herein are methods for the production ofmultiple hybridoma cell lines that produce antibodies specific to IGF-I/II. Further,provided herein are characterization of the antibodies produced by such cell lines,including nucleotide and amino acid sequence analyses of the heavy and light chains ofsuch antibodies.

[0115] Alternatively, instead of being fused to myeloma cells to generatehybridomas, B cells can be directly assayed. For example, CD 19+ B cells can be isolatedfrom hyperimmune XenoMouse® mice and allowed to proliferate and differentiate intoantibody-secreting plasma cells. Antibodies from the cell supernatants are then screened byELISA for reactivity against the IGF-I/II immunogen. The supernatants might also bescreened for immunoreactivity against fragments of IGF-I/II to further map the differentantibodies for binding to domains of functional interest on IGF-I/II. The antibodies mayalso be screened against other related human chemokines and against the rat, the mouse,and non-human primate, such as cynomolgus monkey, orthologues of IGF-I/II, the last todetermine species cross-reactivity. B cells from wells containing antibodies of interest maybe immortalized by various methods including fusion to make hybridomas either fromindividual or from pooled wells, or by infection with EBV or transfection by known 30 P/l 5804/125647/596826/1 191718/2 immortalizing genes and then plating in suitable medium. Alternatively, single plasmacells secreting antibodies with the desired specificities are then isolated using an IGF-I/II-specific hemolytic plaque assay (Babcook et al., Proc. Natl. Acad. Sci. USA 93:7843-48(1996)). Cells targeted for lysis are preferably sheep red blood cells (SRBCs) coated withthe IGF-I/II antigen.

[0116] In the presence of a B-cell culture containing plasma cells secreting theimmunoglobulin of interest and complement, the formation of a plaque indicates specificIGF-I/II-mediated lysis of the sheep red blood cells surrounding the plasma cell of interest.The single antigen-specific plasma cell in the center of the plaque can be isolated and thegenetic information that encodes the specificity of the antibody is isolated from the singleplasma cell. Using reverse-transcription followed by PCR (RT-PCR), the DNA encodingthe heavy and light chain variable regions of the antibody can be cloned. Such clonedDNA can then be further inserted into a suitable expression vector, preferably a vectorcassette such as a pcDNA, more preferably such a pcDNA vector containing the constantdomains of immunglobulin heavy and light chain. The generated vector can then betransfected into host cells, e.g., HEK293 cells, CHO cells, and cultured in conventionalnutrient media modified as appropriate for inducing transcription, selecting transformants,or amplifying the genes encoding the desired sequences.

[0117] In general, antibodies produced by the fused hybridomas were humanIgG2 heavy chains with fully human kappa or lambda light chains. Antibodies describedherein possess human IgG4 heavy chains as well as IgG2 heavy chains. Antibodies canalso be of other human isotypes, including IgGl. The antibodies possessed high affinities,typically possessing a Kd of from about 10" through about 10' M or below, whenmeasured by solid phase and solution phase techniques. Antibodies possessing a KD of atleast 10'11 M are preferred to inhibit the activity of IGF-I/II.

[0118] As will be appreciated, anti-IGF-I/II antibodies can be expressedin cell lines other than hybridoma cell lines. Sequences encoding particular antibodies canbe used to transform a suitable mammalian host cell. Transformation can be by anyknown method for introducing polynucleotides into a host cell, including, forexample packaging the polynucleotide in a virus (or into a viral vector) and transducinga host cell with the virus (or vector) or by transfection procedures known in theart, as exemplified by U.S. Patent Nos. 4,399,216, 4,912,040, 4,740,461, and 4,959,455. The transformation procedure used depends upon the host to betransformed. Methods for introducing heterologous polynucleotides 31 P/15804/125647/596826/l WO 2007/070432 PCT/US2006/047059 into mammalian cells are well known in the art and include dextran-mediatedtransfection, calcium phosphate precipitation, polybrene mediated transfection, protoplastfusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, and directmicroinjection of the DNA into nuclei.

[0119] Mammalian cell lines available as hosts for expression are well knownin the art and include many immortalized cell lines available from the American TypeCulture Collection (ATCC), including but not limited to Chinese hamster ovary (CHO)cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), humanhepatocellular carcinoma cells (e.g., Hep G2), human epithelial kidney 293 cells, and anumber of other cell lines. Cell lines of particular preference are selected throughdetermining which cell lines have high expression levels and produce antibodies withconstitutive IGF-I/II binding properties.

[0120] Anti-IGF-I/II antibodies are useful in the detection of IGF-I/II inpatient samples and accordingly are useful as diagnostics for disease states as describedherein. In addition, based on their ability to significantly neutralize IGF-I/II activity (asdemonstrated in the Examples below), anti-IGF-I/II antibodies have therapeutic effects intreating symptoms and conditions resulting from IGF-I/II expression. In specificembodiments, the antibodies and methods herein relate to the treatment of symptomsresulting from IGF-I/II induced cell proliferation. Further embodiments involve using theantibodies and methods described herein to treat diseases including neoplastic diseases,such as, melanoma, non-small cell lung cancer, glioma, hepatocellular (liver) carcinoma,thyroid tumor, gastric (stomach) cancer, prostrate cancer, breast cancer, ovarian cancer,bladder cancer, lung cancer, glioblastoma, endometrial cancer, kidney cancer, coloncancer, gynecologic tumors, head and neck cancer, esophageal cancer, and pancreaticcancer. Other non-neoplastic disease conditions may include acromegaly and gigantism,psoriasis, osteoporosis, atherosclerosis and smooth muscle restenosis of blood vessels, aswell as diabetes.

Therapeutic Administration and Formulations [0121] Embodiments of the invention include sterile pharmaceutical

formulations of anti-IGF-I/II antibodies that are useful as treatments for diseases. Suchformulations would inhibit the binding of IGF-I/II to its receptor IGF-IR, therebyeffectively treating pathological conditions where, for example, serum or tissue IGF-I/II 32 WO 2007/070432 PCT/US2006/047059 is abnormally elevated. Anti-IGF-1/Π antibodies preferably possess adequate affinity topotently neutralize IGF-I/II, and preferably have an adequate duration of action to allowfor infrequent dosing in humans. A prolonged duration of action will allow for lessfrequent and more convenient dosing schedules by alternate parenteral routes such assubcutaneous or intramuscular injection.

[0122] Sterile formulations can be created, for example, by filtration throughsterile filtration membranes, prior to or following lyophilization and reconstitution of theantibody. The antibody ordinarily will be stored in lyophilized form or in solution.Therapeutic antibody compositions generally are placed into a container having a sterileaccess port, for example, an intravenous solution bag or vial having an adapter that allowsretrieval of the formulation, such as a stopper pierceable by a hypodermic injectionneedle.

[0123] The route of antibody administration is in accord with known methods,e.g., injection or infusion by intravenous, intraperitoneal, intracerebral, intramuscular,intraocular, intraarterial, intrathecal, inhalation or intralesional routes, or by sustainedrelease systems as noted below. The antibody is preferably administered continuously byinfusion or by bolus injection.

[0124] An effective amount of antibody to be employed therapeutically willdepend, for example, upon the therapeutic objectives, the route of administration, and thecondition of the patient. Accordingly, it is preferred that the therapist titer the dosage andmodify the route of administration as required to obtain the optimal therapeutic effect.Typically, the clinician will administer antibody until a dosage is reached that achievesthe desired effect. The progress of this therapy is easily monitored by conventionalassays or by the assays described herein.

[0125] Antibodies, as described herein, can be prepared in a mixture with apharmaceutically acceptable carrier. This therapeutic composition can be administeredintravenously or through the nose or lung, preferably as a liquid or powder aerosol(lyophilized). The composition may also be administered parenterally or subcutaneouslyas desired. When administered systemically, the therapeutic composition should besterile, pyrogen-free and in a parenterally acceptable solution having due regard for pH,isotonicity, and stability. These conditions are known to those skilled in the art. Briefly,dosage formulations of the compounds described herein are prepared for storage oradministration by mixing the compound having the desired degree of purity withphysiologically acceptable carriers, excipients, or stabilizers. Such materials are non- 33 WO 2007/070432 PCT/US2006/047059 toxic to the recipients at the dosages and concentrations employed, and include bufferssuch as TRIS HCI, phosphate, citrate, acetate and other organic acid salts; antioxidantssuch as ascorbic acid; low molecular weight (less than about ten residues) peptides suchas polyarginine, proteins, such as serum albumin, gelatin, or immunoglobulins;hydrophilic polymers such as polyvinylpyrrolidinone; amino acids such as glycine,glutamic acid, aspartic acid, or arginine; monosaccharides, disaccharides, and othercarbohydrates including cellulose or its derivatives, glucose, mannose, or dextrins;chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; counterionssuch as sodium and/or nonionic surfactants such as TWEEN, PLURONICS orpolyethyleneglycol.

[0126] Sterile compositions for injection can be formulated according toconventional phannaceutical practice as described in Remington: The Science andPractice of Pharmacy (20th ed, Lippincott Williams & Wilkens Publishers (2003)). Forexample, dissolution or suspension of the active compound in a vehicle such as water ornaturally occurring vegetable oil like sesame, peanut, or cottonseed oil or a synthetic fattyvehicle like ethyl oleate or the like may be desired. Buffers, preservatives, antioxidantsand the like can be incorporated according to accepted pharmaceutical practice.

[0127] Suitable examples of sustained-release preparations includesemipermeable matrices of solid hydrophobic polymers containing the polypeptide, whichmatrices are in the form of shaped articles, films or microcapsules. Examples ofsustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) as described by Langer et al., J. Biomed Mater. Res., (1981) 15:167-277and Langer, Chem. Tech., (1982) 12:98-105, or poly(vinylalcohol)), polylactides (U.S.Pat. No. 3,773,919, EP 58,481), copolymers of L-glutamic acid and gamma ethyl-L-glutamate (Sidman et al., Biopolymers, (1983) 22:547-556), non-degradable ethylene-vinyl acetate (Langer et al., supra), degradable lactic acid-glycolic acid copolymers suchas the LUPRON Depot™ (injectable microspheres composed of lactic acid-glycolic acidcopolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid (EP 133,988).

[0128] While polymers such as ethylene-vinyl acetate and lactic acid-glycolicacid enable release of molecules for over 100 days, certain hydrogels release proteins forshorter time periods. When encapsulated proteins remain in the body for a long time,they may denature or aggregate as a result of exposure to moisture at 37°C, resulting in aloss of biological activity and possible changes in immunogenicity. Rational strategies 34 WO 2007/070432 PCT/TJS2006/047059 can be devised for protein stabilization depending on the mechanism involved. Forexample, if the aggregation mechanism is discovered to be intermolecular S-S bondformation through disulfide interchange, stabilization may be achieved by modifyingsulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content,using appropriate additives, and developing specific polymer matrix compositions.

[0129] Sustained-released compositions also include preparations of crystalsof the antibody suspended in suitable formulations capable of maintaining crystals insuspension. These preparations when injected subcutaneously or intraperitonealy canproduce a sustained release effect. Other compositions also include liposomal lyentrapped antibodies. Liposomes containing such antibodies are prepared by methodsknown per se: U.S. Pat. No. DE 3,218,121; Epstein et al., Proc. Natl. Acad. Sci. USA,(1985) 82:3688-3692; Hwang et al., Proc. Natl. Acad. Sci. USA, (1980) 77:4030-4034;EP 52,322; EP 36,676; EP 88,046; EP 143,949; 142,641; Japanese patent application 83-118008; U.S. Pat. Nos. 4,485,045 and 4,544,545; and EP 102,324.

[0130] The dosage of the antibody formulation for a given patient will bedetermined by the attending physician taking into consideration various factors known tomodify the action of drugs including severity and type of disease, body weight, sex, diet,time and route of administration, other medications and other relevant clinical factors.Therapeutically effective dosages may be determined by either in vitro or in vivomethods.

[0131] An effective amount of the antibodies, described herein, to beemployed therapeutically will depend, for example, upon the therapeutic objectives, theroute of administration, and the condition of the patient. Accordingly, it is preferred forthe therapist to titer the dosage and modify the route of administration as required toobtain the optimal therapeutic effect. A typical daily dosage might range from about0.001 mg/kg to up to lOOmg/kg or more, depending on the factors mentioned above.Typically, the clinician will administer the therapeutic antibody until a dosage is reachedthat achieves the desired effect. The progress of this therapy is easily monitored byconventional assays or as described herein.

[0132] It will be appreciated that administration of therapeutic entities inaccordance with the compositions and methods herein will be administered with suitablecarriers, excipients, and other agents that are incorporated into formulations to provideimproved transfer, delivery, tolerance, and the like. These formulations include, forexample, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic) 35 WO 2007/070432 PCT/US2006/047059 containing vesicles (such as Lipofectin™), DNA conjugates, anhydrous absorptionpastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethyleneglycols of various molecular weights), semi-solid gels, and semi-solid mixturescontaining carbowax. Any of the foregoing mixtures may be appropriate in treatmentsand therapies in accordance with the present invention, provided that the active ingredientin the formulation is not inactivated by the formulation and the formulation isphysiologically compatible and tolerable with the route of administration. See alsoBaldrick P. “Pharmaceutical excipient development: the need for preclinical guidance.”Regul. Toxicol. Pharmacol. 32(2):210-8 (2000), Wang W. “Lyophilization anddevelopment of solid protein pharmaceuticals.” Int. J. Pharm. 203(1-2):1-60 (2000),Charman WN “Lipids, lipophilic drugs, and oral drug delivery-some emerging concepts.”J Pharm Sci .89(8):967-78 (2000), Powell et al. “Compendium of excipients forparenteral formulations” PDA J Pharm Sci Technol. 52:238-311 (1998) and the citationstherein for additional information related to formulations, excipients and carriers wellknown to pharmaceutical chemists.

Design and Generation of Other Therapeutics [0133] In accordance with the present invention and based on the activity ofthe antibodies that are produced and characterized herein with respect to IGF-I/II, thedesign of other therapeutic modalities is facilitated and disclosed to one of skill in the art.Such modalities include, without limitation, advanced antibody therapeutics, such asbispecific antibodies, immunotoxins, radiolabeled therapeutics, and single antibody Vdomains, antibody-like binding agent based on other than V region scaffolds, generationof peptide therapeutics, gene therapies, particularly intrabodies, antisense therapeutics,and small molecules.

[0134] In connection with the generation of advanced antibody therapeutics,where complement fixation is a desirable attribute, it can be possible to sidestep thedependence on complement for cell killing through the use of bispecifics, immunotoxins,or radiolabels, for example.

[0135] For example, bispecific antibodies can be generated that comprise (i)two antibodies, one with a specificity to IGF-I/II and another to a second molecule, thatare conjugated together, (ii) a single antibody that has one chain specific to IGF-I/II and asecond chain specific to a second molecule, or (iii) a single chain antibody that hasspecificity to both IGF-I/II and the other molecule. Such bispecific antibodies can be 36 WO 2007/070432 PCT/US2006/047059 generated using techniques that are well known; for example, in connection with (i) and (ii) see e.g., Fanger et al. Immunol Methods 4:72-81 (1994) and Wright and Harris, supra.and in connection with (iii) see e.g., Traunecker et al. Int. J. Cancer (Suppl.) 7:51-52(1992). In each case, the second specificity can be made as desired. For example, thesecond specificity can be made to the heavy chain activation receptors, including, withoutlimitation, CD16 or CD64 (see e.g., Deo et al. 18:127 (1997)) or CD89 (see e.g., Valeriuset al. Blood 90:4485-4492 (1997)).

[0136] Antibodies can also be modified to act as immunotoxins utilizingtechniques that are well known in the art. See e.g., Vitetta Immunol Today 14:252 (1993).See also U.S. Patent No. 5,194,594. In connection with the preparation of radiolabeledantibodies, such modified antibodies can also be readily prepared utilizing techniques thatare well known in the art. See e.g., Junghans et al. in Cancer Chemotherapy andBiotherapy 655-686 (2d edition, Chafner and Longo, eds., Lippincott Raven (1996)). Seealso U.S. Patent Nos. 4,681,581, 4,735,210, 5,101,827, 5,102,990 (RE 35,500),5,648,471, and 5,697,902. Each of immunotoxins and radiolabeled molecules would belikely to kill cells expressing the desired multimeric enzyme subunit oligomerizationdomain. In some embodiments, a pharmaceutical composition comprising an effectiveamount of the antibody in association with a pharmaceutically acceptable carrier ordiluent is provided.

[0137] In some embodiments, an anti-IGF-I/II antibody is linked to an agent(e.g., radioisotope, pharmaceutical composition, or a toxin). Preferably, such antibodiescan be used for the treatment of diseases, such diseases can relate to cells expressing IGF-I/II or cells overexpressing IGF-I/II. For example, it is contemplated that the drugpossesses the pharmaceutical property selected from the group of antimitotic, alkylating,antimetabolite, antiangiogenic, apoptotic, alkaloid, COX-2, and antibiotic agents andcombinations thereof. The drug can be selected from the group of nitrogen mustards,ethylenimine derivatives, alkyl sulfonates, nitrosoureas, triazenes, folic acid analogs,anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs,antimetabolites, antibiotics, enzymes, epipodophyllotoxins, platinum coordinationcomplexes, vinca alkaloids, substituted ureas, methyl hydrazine derivatives,adrenocortical suppressants, antagonists, endostatin, taxols, camptothecins, oxaliplatin,doxorubicins and their analogs, and a combination thereof.

[0138] Examples of toxins further include gelonin, Pseudomonas exotoxin(PE), PE40, PE38, diphtheria toxin, ricin, ricin, abrin, alpha toxin, saporin, ribonuclease 37 WO 2007/070432 PCT/US2006/047059 (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin,Pseudomonas endotoxin, as well as derivatives, combinations and modifications thereof.

[0139] Examples of radioisotopes include gamma-emitters, positron-emitters,and x-ray emitters that can be used for localization and/or therapy, and beta-emitters andalpha-emitters that can be used for therapy. The radioisotopes described previously asuseful for diagnostics, prognostics and staging are also useful for therapeutics. Non-limiting examples of anti-cancer or anti-leukemia agents include anthracyclines such asdoxorubicin (adriamycin), daunorubicin (daunomycin), idarubicin, detorubicin,carminomycin, epirubicin, esorubicin, and morpholino and substituted derivatives,combinations and modifications thereof. Exemplary pharmaceutical agents include cis-platinum, taxol, calicheamicin, vincristine, cytarabine (Ara-C), cyclophosphamide,prednisone, daunorubicin, idarubicin, fludarabine, chlorambucil, interferon alpha,hydroxyurea, temozolomide, thalidomide, and bleomycin, and derivatives, combinationsand modifications thereof. Preferably, the anti-cancer or anti-leukemia is doxorubicin,morpholinodoxorubicin, or morpholinodaunorubicin.

[0140] As will be appreciated by one of skill in the art, in the aboveembodiments, while affinity values can be important, other factors can be as important ormore so, depending upon the particular function of the antibody. For example, for animmunotoxin (toxin associated with an antibody), the act of binding of the antibody to thetarget can be useful; however, in some embodiments, it is the internalization of the toxininto the cell that is the desired end result. As such, antibodies with a high percentinternalization can be desirable in these situations. Thus, in one embodiment, antibodieswith a high efficiency in internalization are contemplated. A high efficiency ofinternalization can be measured as a percent internalized antibody, and can be from a lowvalue to 100%. For example, in varying embodiments, 0.1-5, 5-10, 10-20, 20-30, 30-40,40-45, 45-50, 50-60, 60-70, 70-80, 80-90, 90-99, and 99-100% can be a high efficiency.As will be appreciated by one of skill in the art, the desirable efficiency can be differentin different embodiments, depending upon, for example, the associated agent, the amountof antibody that can be administered to an area, the side effects of the antibody-agentcomplex, the type (e.g., cancer type) and severity of the problem to be treated.

[0141] In other embodiments, the antibodies disclosed herein provide an assaykit for the detection of IGF-I/II expression in mammalian tissues or cells in order toscreen for a disease or disorder associated with changes in expression of IGF-I/II. The kit 38 WO 2007/070432 PCT/US2006/047059 comprises an antibody that binds IGF-I/II and means for indicating the reaction of theantibody with the antigen, if present.

[0142] In some embodiments, an article of manufacture is providedcomprising a container, comprising a composition containing an anti-IGF-I/II antibody,and a package insert or label indicating that the composition can be used to treat diseasemediated by IGF-I/II expression. Preferably a mammal, and more preferably, a human,receives the anti-IGF-I/II antibody.

Combinations [0143] The anti-IGF-Ι/Π antibodies defined hereinbefore may be applied as asole therapy or may involve, in addition to the compound of the invention, conventionalsurgery or radiotherapy or chemotherapy. Such chemotherapy may include one or moreof the following categories of anti-tumour agents :- (i) antiproliferative/antineoplastic drugs and combinations thereof, as usedin medical oncology, such as alkylating agents (for example cis-platin, carboplatin,cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan andnitrosoureas); antimetabolites (for example antifolates such as fluoropyrimidines like5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside andhydroxyurea; antitumour antibiotics (for example anthracyclines like adriamycin,bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycinand mithramycin); antimitotic agents (for example vinca alkaloids like vincristine,vinblastine, vindesine and vinorelbine and taxoids like taxol and taxotere); andtopoisomerase inhibitors (for example epipodophyllotoxins like etoposide and teniposide,amsacrine, topotecan and camptothecin); (ii) cytostatic agents such as antioestrogens (for example tamoxifen,toremifene, raloxifene, droloxifene and iodoxyfene), oestrogen receptor down regulators(for example fulvestrant), antiandrogens (for example bicalutamide, flutamide, nilutamideand cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin,leuprorelin and buserelin), progestogens (for example megestrol acetate), aromataseinhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitorsof 5a-reductase such as finasteride; 39 WO 2007/070432 PCT/US2006/047059 (iii) agents which inhibit cancer cell invasion (for examplemetalloproteinase inhibitors like marimastat and inhibitors of urokinase plasminogenactivator receptor function); (iv) inhibitors of growth factor function, for example such inhibitors include growth factor antibodies, growth factor receptor antibodies (for example theanti-erbb2 antibody trastuzumab [Herceptin™] and the anti-erbbl antibody cetuximab[C225]) , famesyl transferase inhibitors, MEK inhibitors, tyrosine kinase inhibitors andserine/threonine kinase inhibitors, for example inhibitors of the epidermal growth factorfamily (for example EGFR family tyrosine kinase inhibitors such as W-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, AZD1839), JV-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-aniine (erlotinib,OSI-774) and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3- morpholinopropoxy)quinazolin-4-amine (CI 1033)), for example inhibitors of theplatelet-derived growth factor family and for example inhibitors of the hepatocyte growthfactor family; (v) antiangiogenic agents such as those which inhibit the effects ofvascular endothelial growth factor, (for example the anti-vascular endothelial cell growthfactor antibody bevacizumab [Avastin™], compounds such as those disclosed inInternational Patent Applications WO 97/22596, WO 97/30035, WO 97/32856 and WO98/13354) and compounds that work by other mechanisms (for example linomide,inhibitors of integrin ανβ3 function; angiostatin and inhibitors of the action ofangiopoietins e.g angiopoietin 1 and angiopoietin 2); (vi) vascular damaging agents such as Combretastatin A4 and compounds disclosed in International Patent Applications WO 99/02166, WO00/40529, WO 00/41669, WOO1/92224, W002/04434 and W002/08213; (vii) antisense therapies, for example those which are directed to the targets listedabove, such as ISIS 2503, an anti-ras antisense; (viii) gene therapy approaches, including for example approaches to replaceaberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT(gene-directed enzyme pro-drug therapy) approaches such as those using cytosinedeaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches toincrease patient tolerance to chemotherapy or radiotherapy such as multi-drug resistancegene therapy; 40 WO 2007/070432 PCT/US2006/047059 (ix) immunotherapy approaches, including for example ex-vivo and in-vivoapproaches to increase the immunogenicity of patient tumour cells, such as transfectionwith cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colonystimulating factor, approaches to decrease T-cell anergy, approaches using transfectedimmune cells such as cytokine-transfected dendritic cells, approaches usingcytokine-transfected tumour cell lines and approaches using anti-idiotypic antibodies; (x) cell cycle inhibitors including for example CDK inhibitiors (eg flavopiridol)and other inhibitors of cell cycle checkpoints (eg checkpoint kinase); inhibitors of aurorakinase and other kinases involved in mitosis and cytokinesis regulation (eg mitotickinesins); and histone deacetylase inhibitors; (xi) endothelin antagonists, including endothelin A antagonists, endothelin Bantagonists and endothelin A and B antagonists; for example ZD4054 and ZD1611 (WO96 40681), atrasentan and YM598; and (xii) biotherapeutic therapeutic approaches for example those which usepeptides or proteins (such as antibodies or soluble external receptor domainconstructions) which either sequest receptor ligands, block ligand binding to receptor ordecrease receptor signalling (e.g. due to enhanced receptor degradation or loweredexpression levels) [0144] Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of the treatment. Suchcombination products employ the compounds of this invention within the dosage rangedescribed hereinbefore and the other pharmaceutically-active agent within its approveddosage range.

EXAMPLES

[0145] The following examples, including the experiments conducted andresults achieved are provided for illustrative purposes only and are not to be construed aslimiting upon the teachings herein. EXAMPLE 1

Immunization and TITERING 41 WO 2007/070432 PCT/US2006/047059

Immunization [0146] Recombinant human IGF-I and IGF-II obtained from R&D Systems,Inc. (Minneapolis, MN Cat. No. 291 -G1 and 292-G2 respectively) were used as antigens.Monoclonal antibodies against IGF-I/II were developed by sequentially immunizingXenoMouse® mice (XenoMouse strains XMG2 and XMG4 (3C-1 strain), Abgenix, Inc.Fremont, CA). XenoMouse animals were immunized via footpad route for all injections.The total volume of each injection was 50 μΐ per mouse, 25 μΐ per footpad. A total of ten(10) mice were immunized in each group. Each injection was with 10 pg per mouse ofIGF-I or IGF-II alone or conjugated to Keyhole Limpet Hemocyanin (KLH) antigen as acarrier, as detailed in Table 2. The first injection was made up in Dulbecco’s PBS(DPBS) and admixed 1:1 v/v with Titermax Gold Adjuvant (SIGMA Cat. #T2684, lot#K1599). A total of 8 to II additional boosts were then administered over a period of 27to 38 days, admixed with 25 pg of Adju-Phos (aluminum phosphate gel, Catalog # 1452-250, batch #8937, HCI Biosector) and 10 pg CpG (15 pi of ImmunEasy Mouse Adjuvant,catalog # 303101; lot #11553042; Qiagen) per mouse, followed by a final boost of 10 pgof antigen in pyrogen-free DPBS, without adjuvant. For combined immunization(animals immunized with both IGF-I and IGF-II), the second antigen was given in the lasttwo (2) boosts.

TABLE 2. IMMUNIZATION SUMMARY

Immunization Group Initial Immunogen Final Immunogen KLH Conjugated Isotype ofMice Fusion Group 1 IGF-1 IGF-1 — IgG2-KX 1 3 IGF-1 IGF-1 — IgG4-KA 1 5 IGF-1 IGF-1 4* IgG2-tcX 1 7 IGF-1 IGF-1 + IgG4-x% 1 2 IGF-2 IGF-2 — IgG2-KX 2 4 IGF-2 IGF-2 - IgG4-xX 2 6 IGF-2 IGF-2 + IgG2-KX 2 8 IGF-2 IGF-2 + IgG4-KX 2 9 IGF-1 IGF-2 - IgG2-K% 3 11 IGF-1 IGF-2 — IgG4-xX 3 13 IGF-I IGF-2 + IgG2-Kl 3 15 IGF-1 IGF-2 + IgG4-KX 3 10 IGF-2 IGF-1 — IgG2-K?l 4 12 IGF-2 IGF-1 — IgG4-KX 4 42 WO 2007/070432 PCT/US2006/047059 14 IGF-2 IGF-1 + IgG2-xX 4 16 IGF-2 IGF-1 + IgG4-KX 4 EXAMPLE 2

RECOVERY OF LYMPHOCYTES, B-CELL ISOLATIONS, FUSIONS ANDGENERATION OF HYBRIDOMAS

[0147] Immunized mice were sacrificed by cervical dislocation, and thedraining lymph nodes harvested and pooled from each cohort. The lymphoid cells weredissociated by grinding in DMEM to release the cells from the tissues and the cells weresuspended in DMEM. The cells were counted, and 0.9 ml DMEM per 100 millionlymphocytes added to the cell pellet to resuspend the cells gently but completely. Using100 μΐ of CD90+ magnetic beads per 100 million cells, the cells were labeled byincubating the cells with the magnetic beads at 4°C for 15 minutes. The magneticallylabeled cell suspension containing up to 108 positive cells (or up to 2xl09 total cells) wasloaded onto a LS+ column and the column washed with DMEM. The total effluent wascollected as the CD90-negative fraction (most of these cells were expected to be B cells).

[0148] The fusion was performed by mixing washed enriched B cells fromabove and nonsecretory myeloma P3X63Ag8.653 cells purchased from ATCC, cat.# CRL1580 (Kearney et al, J. Immunol. 123, 1979, 1548-1550) at a ratio of 1:1. The cellmixture was gently pelleted by centrifugation at 800 x g. After complete removal of thesupernatant, the cells were treated with 2-4 mL of Pronase solution (CalBiochem, cat. #53702; 0.5 mg/ml in PBS) for no more than 2 minutes. Then 3-5 ml of FBS was added tostop the enzyme activity and the suspension was adjusted to 40 ml total volume usingelectro cell fusion solution, (ECFS, 0.3M Sucrose, Sigma, Cat# S7903, 0.1 mMMagnesium Acetate, Sigma, Cat# M2545, O.lmM Calcium Acetate, Sigma, Cat# C4705).The supernatant was removed after centrifugation and the cells were resuspended in 40ml ECFS. This wash step was repeated and the cells again were resuspended in ECFS to aconcentration of 2x106 cells/ml.

[0149] Electro-cell fusion was performed using a fusion generator (modelECM2001, Genetronic, Inc., San Diego, CA). The fusion chamber size used was 2.0 ml,using the following instrument settings: [0150] Alignment condition: voltage: 50 V, time: 50 sec.

[0151] Membrane breaking at: voltage: 3000 V, time: 30 psec 43 WO 2007/070432 PCT/US2006/047059 [0152] Post-fusion holding time: 3 sec [0153] After ECF, the cell suspensions were carefully removed from thefusion chamber under sterile conditions and transferred into a sterile tube containing thesame volume of Hybridoma Culture Medium (DMEM, JRH Biosciences), 15 % FBS(Hyclone), supplemented with L-glutamine, pen/strep, OPI (oxaloacetate, pyruvate,bovine insulin) (all from Sigma) and IL-6 (Boehringer Mannheim). The cells wereincubated for 15-30 minutes at 37°C, and then centrifuged at 400 x g (1000 rpm) for fiveminutes. The cells were gently resuspended in a small volume of Hybridoma SelectionMedium (Hybridoma Culture Medium supplemented with 0.5x HA (Sigma, cat. #A9666)), and the volume adjusted appropriately with more Hybridoma SelectionMedium, based on a final plating of 5xl06 B cells total per 96-well plate and 200 μΐ perwell. The cells were mixed gently and pipetted into 96-well plates and allowed to grow.On day 7 or 10, one-half the medium was removed, and the cells re-fed with HybridomaSelection Medium. EXAMPLE 3

SELECTION OF CANDIDATE ANTIBODIES BY ELISA

[0154] After 14 days of culture, hybridoma supernatants were screened forIGF-I/II-specific monoclonal antibodies. The ELISA plates (Fisher, Cat. No. 12-565-136) were coated with 50 μΐ/well of human IGF-I or IGF-II (2 μg/ml) in Coating Buffer(0.1 M Carbonate Buffer, pH 9.6, NaHCO3 8.4 g/L), then incubated at 4°C overnight.After incubation, the plates were washed with Washing Buffer (0.05% Tween 20 in PBS)3 times. 200 μΐ/well Blocking Buffer (0.5% BSA, 0.1% Tween 20, 0.01% Thimerosal inlx PBS) were added and the plates incubated at room temperature for 1 hour. Afterincubation, the plates were washed with Washing Buffer three times. 50 μΐ/well ofhybridoma supernatants, and positive and negative controls were added and the platesincubated at room temperature for 2 hours.

[0155] After incubation, the plates were washed three times with WashingBuffer. 100 μΙ/well of detection antibody goat anti-huIgGFc-HRP (Caltag, Cat.No. Hl 0507), was added and the plates incubated at room temperature for 1 hour. In asecondary screen, the positives in first screening were screened in two sets, one forhuman IgG (heavy chain) detection and the other for human Ig kappa light chaindetection (goat anti-hlg kappa-HRP (Southern Biotechnology, Cat. No. 2060-05) in orderto demonstrate fully human composition for both IgG and Ig kappa. After incubation, the 44 WO 2007/070432 PCT/US2006/047059 plates were washed three times with Washing Buffer. 100 μΙ/well of TMB (BioFX Lab.Cat. No. TMSK-0100-01) were added and the plates allowed to develop for about 10minutes (until negative control wells barely started to show color). 50 μΙ/well stopsolution (TMB Stop Solution, (BioFX Lab. Cat. No. STPR-0100-01) was then added andthe plates read on an ELISA plate reader at 450nm. As indicated in Table 3, there were atotal of 1,233 wells containing antibodies against IGF-I and -II.

[0156] All antibodies that bound in the ELISA assay were counter screenedfor binding to insulin by ELISA in order to exclude those that cross-reacted with insulin.The ELISA plates (Fisher, Cat. No. 12-565-136) were coated with 50 μΙ/well ofrecombinant insulin (concentration: lgg/ml; Sigma, catalog # 12643) in Coating Buffer(0.1 M Carbonate Buffer, pH 9.6, NaHCO3 8.4 g/L), then incubated at 4°C overnight. Asdetailed in Table 3, a total of 1,122 antibodies from the original 1233 antibodies did crossreact with insulin.

TABLE 3. SCREENING SUMMARY

Confirmation Screen Fn # Mouse strain Immunogen Targetwith hGdetection Target with hK+hL detection IGF-1(+) &IGF-H(+) 1GF-I(+) & IGF- 11(+) andhu- Insulin (-) 1 G2KL IGF-I-KLH IGF-II IGF-I 36 28 2 G4KL IGF-I-KLH IGF-II IGF-I 65 55 3 G2KL IGF-II orIGF-II-KLH IGF-I IGF-II 168 150 4 G4KL IGF-Π orIGF-II-KLH IGF-I IGF-Π 197 194 5 G2KL IGF-I/-II- KLH IGF-II IGF-II 54 50 6 G4KL IGF-I/-II- KLH IGF-II IGF-II 101 86 7 G2KL IGF-II/-I- KLH IGF-II IGF-II 294 271 8 G4KL IGF-II/-I orIGF-II/-I-KLH IGF-II IGF-II 318 288 Total FI to F4 Total F5 to F8 Total 466 427 767 695 1,233 1,122 45 WO 2007/070432 PCT/US2006/047059 [0157] Finally, the antibodies that were selected in the counter-screen werethen tested by ELISA to confirm binding to mouse IGF-I and IGF-II proteins. A total of683 hybridoma lines .were identified that have cross-reactivity with mouse IGF-I/II.Accordingly, these hybridoma lines expressed antibodies that bound to human IGF-I,human IGF-II, mouse IGF-I and mouse IGF-II, but did not bind to human insulin. EXAMPLE 4

INHIBITION OF IGF-I AND IGF-II BINDING TO IGF-IR

[0158] The purpose of this study was to screen the 683 anti-IGF-I/II humanIgG2 and IgG4 antibodies at the hybridoma supernatant stage for neutralizing activity, asdetermined by inhibition of IGF-I and IGF-II binding to the IGF-IR receptor. Thus, areceptor/ligand binding assay was performed with NIH3T3 cells that overexpress thehuman IGF-IR receptor, as described below.

[0159] Briefly, multi-screen filter plates (Multiscreen 0.65 μΜ 96-wellPVDF, Millipore, Cat. No. MADV NOB 10) were blocked with blocking buffer (PBScontaining 10%BSA with 0.02%NaN3) at 200pL/well overnight at 4°C. [125I]-labeled IGF(Amersham Life Sciences Cat No. IM172 (IGF-I) or IM238 (IGF-II)) at 100pCi/ml and50nM was diluted to the appropriate concentration (70pM final for IGF-I and 200pMfinal for IGF-II) in binding buffer (PBS containing 2%BSA with 0.02% NaNa). Theblocking buffer-coated filter plate was washed once with 200pL PBS, and 50pL anti-IGF-I/II Ab supernatants (diluted in binding buffer to 25% final volume) were preincubatedwith 25 pL of [I25I]-IGF in the MultiScreen plate for 30-60 minutes on ice. SubconfluentNIH3T3 mouse fibroblasts stably expressing hIGF-IR (obtained from AstraZeneca) wereharvested with trypsin and resuspended in cold binding buffer at 6xl06/mI, and 25pL ofcells were added to the plate for a two-hour incubation on ice. The plate was washed fourtimes with 200pL cold PBS and dried overnight. Twenty-five pL/well of scintillant(SuperMix cocktail, Wallac/Perkin Elmer Cat No. 1200-439) was added and the plateswere read using a Microbeta Trilux reader (Wallac).

[0160] The following controls were used per screening plate: no antibody(total IGF bound), control neutralizing anti-IGF-I (#05-172, Upstate) or anti-IGF-II(#MAB292, R&D Systems) mAbs at 50pg/ml (non-specific background) and 0.075 to0.5pg/ml (approximate EC50 values of the neutralizing antibodies), and isotype-matchedcontrol human IgG2 (PK16.3.1, Abgenix, lot #360-154) or IgG4 (108.2.1, Abgenix, 46 WO 2007/070432 PCT/US2006/047059 lot#718-53A) mAbs at a concentration of 0.5pg/ml (approximate EC50 value ofneutralizing antibodies). An additional titration of control neutralizing antibodies andisotype-matched control human antibodies was added to one plate per screening assay(1/10 serial dilution from 50ug/ml (333.3nM)). All controls with or without antibodieswere prepared in binding buffer supplemented with anti-KLH human IgG2 or IgG4exhaust supernatant at 25% final volume.

[0161] The percentage of inhibition was determined as follows:

% Inhibition^ ([(Mean CPM Total 12SI-IGF bound)- (Mean CPM 125I-IGF boundin the presence of antibody)] / [(Mean CPM Total ,25I-IGF bound)-(Mean CPM l25I-IGFbound in the presence of an excess of control neutralizing antibody*)]) xlOO

[0162] * the non-specific background was determined as CPM of cells with an excess of control neutralizing anti-IGF Ab (50ug/ml, 333.3nM), which was found to beequivalent to an excess of cold IGF (less than or equal to 10% of total CPM) [0163] The anti-IGF-I/II supernatant screening was split by isotype because ofradiolabeled ligand availability issues. As shown in Table 4, supernatants from the anti-IGF-I/II antibodies with an IgG2 isotype (293 total) were first screened againstradiolabeled IGF-I. A cut-off at 40% inhibition was initially applied to this screening (i.e.hybridoma lines inhibiting at 40% and above were selected), and 111 hits were selectedfor subsequent screening against IGF-Π. Of the 111 hits, a total of 91 lines were found toinhibit IGF-II binding to its receptor with a 50% cut-off. A total of 71 final hits wereselected by taking supernatants that neutralized 50% of both IGF-I and IGF-Π activity.

[0164] All the supernatants expressing IgG4 isotypes (390 total) were initiallyscreened against radiolabeled IGF-II, and 232 hits with a cut-off at 50% inhibition weresubsequently screened against IGF-I. A total of 90 lines were able to inhibit IGF-Ibinding to its receptor with a 50% cut-off. After combining the hits for IgG2 (71) andIgG4 (90), a total of 161 lines were obtained which inhibited IGF-I and IGF-II by 50% ormore.

[0165] In conclusion, from the 683 original supernatants, 343 (111 IgG2 and232 IgG4, 50.2%) were selected from the first screening with either IGF-I or IGF-II. Atotal of 161 final hits were obtained (23.6% of original lines), which are able to block 47 WO 2007/070432 PCT/US2006/047059 both IGF-I and IGF-II binding to IGF-IR with an overall cut-off criteria of 50%inhibition. 48 TABLE 4. ANTI-IGF-l/II EXHAUST SUPERNATANT SCREENING SUMMARY (50% CUT-OFF) ESN Activity Fusion # Mouse strain Immunogen hIGF-II with hG+/hlC or hG+/hL+ R/L BindingAssay IGF-I IGF-I+ % Total R/L Binding AssayIGF-IIon IGF- 1+ IGF- I/II+ % Total R/L Binding Assay IGF-Π IGF-II+ % Total R/L BindingAssayIGF-I onIGF-II+ IGF- I/II+ % Total 1 G2KL IGF-I-KLH 14 12 85.7 1 7.1 2 G4 KL IGF-I-KLH 20 4 20.0 4 20.0 3 G2KL IGF-II orIGF-II-KLH 49 13 26.5 4 8.2 4 G4KL IGF-II orIGF-II-KLH 114 86 75.4 36 31.6 5 G2 KL 1GF-I/-II-KLH 32 II 34.4 7 21.9 6 G4KL IGF-I/-II-KLH 47 21 44.7 8 17.0 7 G2KL IGF-II/-I-KLH 198 75 37.9 59 29.8 8 G4 KL IGF-1I/-I orIGF-II/-I-KLH . 209 121 57.9 42 20.1 683 111 16.3 71 10.4 232 34.0 90 13.2 Cut-off 40% IgG2 IgG4 2007/070432 PCT/US2006/047059 WO 2007/070432 PCT/US2006/047059 EXAMPLE 5

HIGH ANTIGEN AND LIMITED ANTIGEN ELISAS

[0166] In order to determine the relative affinities among the 161 hybridoma linesselected in Example 4, as well as the concentration of antibody in the supernatants of eachline, high antigen (HA) and limited antigen (LA) ELISA assays were carried out. In the HAquantitation assay, the high antigen concentration and overnight incubation limit the effect ofantibody affinity, allowing for quantitation of the relative amount of antigen-specificantibody present in each sample. The low antigen concentration in the LA assay limits theeffect of antibody concentration and results in a ranking of antibodies based on their relativeaffinity.

High Antigen Quantitation Assay J0167] ELISA plates were coated with relatively large amounts of either IGF-I orIGF-II antigen (R&D Systems, Inc., Minneapolis, MN Cat. No. 291-Gl and 292-G2respectively) at 500ng/ml (67nM). Antibody-containing hybridoma supernatants weretitrated over a dilution range of 1:50 to 1:12200. A control of a known IGF-specific antibody(R&D Systems, Inc., Minneapolis, MN Cat. No. MAB291 and MAB292 respectively) wasused to define the linear range of the assay. Data within the linear range were then used toderive the relative concentration of the IGF-specific antibody in each titrated sample.

Limited Antigen Assay [0168] Microtiter plates were coated with low concentrations of antigen. Fiftymicroliters (50 pL) of IGF-I or IGF-II at 64, 32, 16, 8, 4, and 2 ng/ml (covering a range of8.5 nM to 0.26 nM) in 1% skim milk / 1X PBS pH 7.4 / 0.5% azide was added to each well.The plate was incubated for 30 minutes.

[0169] Plates were washed four times (4X) with water, and 50pL of hybridomasupernatant containing test antibodies diluted 1:25 in 1% skim milk I IX PBS pH 7.4 / 0.5%azide were added to the wells. Plates were wrapped tightly with plastic wrap or paraffin film,and incubated overnight with shaking at room temperature.

[0170] On the following day, all plates were washed five times (5X) and 50 pL goat anti-Human IgG Fc HRP polyclonal antibody at a concentration of 0.5 ug/ml in 1% 50 WO 2007/070432 PCT/US2006/047059 milk, IX PBS pH 7.4 was added to each well. The plates were incubated for 1 hour at room temperature. (0171] Plates were washed at least five times (5X with tap water). Fiftymicroliters (50) pL of HPR substrate TMB was added to each well, and the plate wereincubated for 30 minutes. The HRP-TMB reaction was stopped by adding 50 pL of 1Mphosphoric acid to each well. Optical density (absorbance) at 450 nm was measured for eachwell of the plate.

Data Analysis [0172] OD values of test antibodies were averaged and the range was calculated.Antibodies with the highest signal and acceptably low standard deviation were selected asantibodies having a higher affinity for the antigen than did a reference antibody.

[0173] An analysis was then made to select top antibodies based on eitherneutralization (Example 4), potency (low antibody concentration as determined by HAELISA and high inhibition of ligand binding), affinity (LA ELISA), or all three criteria.From this analysis, a list of 25 antibodies was generated. A separate analysis based onaverage % inhibition of IGF-1 and -II binding and affinity for both IGF-I and IGF-IIgenerated a second list of 25 antibodies. Sixteen antibodies were common to both lists,resulting in a final list of 40 antibodies. The LA and HA results for these 40 antibodies aresummarized in Table 5. These 40 lines were selected for cloning, of which 33 weresuccessfully cloned.

TABLE 5. RESULTS OF HIGH AND LIMITED ANTIGEN ELISA FOR TOP 40ANTIBODIES IGF1 HA IGF2 HA IGF1 LA IGF2 LA IGF1 LA IGF2 LA Line ID Avg (ug/ml) HA1 Std. Dev HA1 Avg (ug/ml) HA2 Std. Dev HA2 IGF1 2ng/ml IGF2 2ng/ml IGF1 4ng/ml IGF2 4ng/ml 4.121 1.16 0.16 3.56 1.34 0.56 0.53 1.14 0.90 4.141 1.99 0.22 1.99 0.21. 0.57 0.79 1.25 1.52 4.142 4.70 0.21 3.65 0.31 0.78 1.00 1.76 1.78 4.143 1.74 0.17 2.03 0.41 0.60 0.99 1.20 1.91 4.25 1.26 0.23 1.48 0.36 0.71 0.81 1.31 1.54 51 WO 2007/070432 PCT/US2006/047059 IGF1 HA IGF2 HA IGF1 LA IGF2 LA IGF1 LA 1GF2 LA Line ID Avg (ug/ml) HA1 Std. Dev HA1 Avg (ug/ml) HA2 Std. Dev HA2 IGF1 2ng/ml IGF2 2ng/ml IGF1 4ng/ml IGF2 4ng/ml 4.69 7.17 1.16 6.50 0.53 0.80 0.80 1.50 1.54 4.90 1.15 0.12 3.68 0.77 0.58 0.48 1.04 0.89 7.118 10.34 1.26 10.32 1.90 0.81 0.85 1.56 1.44 7.123 13.4 3.2 12.0 2.8 1.0 1.7 1.66 2.58 7.127 7.28 0.44 6.59 1.55 1.19 1.37 2.29 2.40 7.130 4.32 0.32 3.51 1.34 0.64 1.17 1.36 1.98 7.146 12.04 0.98 9.63 0.6 0.2 0.86 0.29 1.58 7.158 9.29 0.49 7.1 0.56 1.71 1.42 3.00 2.46 7.159 16.53 1.83 41.1 0.56 1.65 0.98 2.47 7.160 4.9 0.5 5.1 0.2 1.7 2.2 3.14 3.30 7.175 8.46 0.49 6.21 1.22 0.13 0.34 0.14 0.62 7.202 11.94 1.98 15.24 1.72 1.11 1.68 2.28 2.69 7.212 11.30 1.90 10.86 1.26 0.97 0.93 2.22 1.54 7.215 10.11 2.05 10.94 1.39 1.01 1.09 2.25 1.93 7.23 4.30 0.26 3.99 0.29 0.55 1.22 1.40 2.17 7.234 4.7 1.4 3.1 0.4 0.7 1.4 1.79 2.44 7.251 3 0.41 1.93 0.17 1.09 1.02 1.31 1.53 7.252 8.25 0.51 5.55 1.68 1.22 1.23 2.53 2.09 7.268 7.58 0.42 5.07 0.92 1.47 1.37 3.06 2.42 7.29 12.5 1.24 23.53 4.7 0.18 0.39 0.27 0.49 7.3 13.18 2.12 8.83 0.58 0.81 1.28 2.07 1.96 7.34 12.54 1.99 14.67 3.05 0.21 1.07 0.44 1.84 7.41 3.69 0.19 4.97 0.8 1.02 1.53 2.21 2.32 7.56 14.6 2.0 21.7 3.7 1.3 1.4 2.38 2.46 7.58 17.52 0.01 27.54 6.22 0.21 1.15 0.47 1.82 7.66 6.02 0.81 6.18 0.71 0.49 0.97 1.42 1.53 7.77 8.42 0.18 7.25 1.12 0.64 0.46 1.50 1.00 7.85 22.67 0.68 23.63 0.93 0.1 0.33 0.16 0.51 7.99 7.9 0.2 5.9 1.6 0.9 1.0 1.87 1.65 8.119 1.26 0.00 0.77 0.16 2.37 0.79 3.77 1.36 8.141 5.96 0.50 4.12 0.61 1.80 0.61 3.02 1.08 8.146 4.03 0.45 2.55 0.74 0.97 1.06 2.13 1.98 8.287 4.8 0.1 2.8 0.8 2.2 1.6 3.80 2.91 8.8 2.00 0.17 1.45 0.25 1.72 0.77 3.13 1.46 8.86 3.15 0.19 2.36 0.24 0.92 1.35 1.76 2.18 52 WO 2007/070432 PCT/US2006/047059 EXAMPLE 6

BfNDING OF ANTIBODIES TO IGF-I AND IGF-II BOUND TO IGFBP-3 [0174] IGF-I and -II circulate in serum mostly bound to IGF-binding proteins(IGFBPs). One aim was to identify antibodies that do not recognize IGFs in complex withIGFBPs, in order to avoid in vivo depletion of anti-IGF antibodies. The following assayformat was developed for the characterization of antibodies that recognize IGF-I or IGF-IIwhen these growth factors are complexed with IGFBP-3. Specifically, this assay tested theability of IGF in IGF/anti-IGF antibody complexes to bind IGFBP-3.

Antibody-Mediated Block of Capture of IGF by IGFBP-3 [0175] An assay was developed wherein complexes were pre-formed betweenIGF-I or IGF-II and IGF-specific antibodies from the aforementioned examples. The abilityof these complexes to bind to IGFBP-3 was tested using AlphaScreen assay technology(PerkinElmer). In a 384-well plate, 10 pL 1:20 diluted hybridoma supernatants were mixedwith 10 pL of 3 nM biotinylated IGF-I or IGF-II and incubated at room temperature for 2hours. Streptavidin-coated AlphaScreen donor beads and IGFBP-3-coupled AlphaScreenacceptor beads (10 uL of a mixture, for a 1/60 final dilution of the hybridoma supernatants)were added, and the incubation was continued for another hour. Samples were then read in aPackard Fusion plate reader.

[0176] Three commercially available anti-IGF monoclonal antibodies M23 (CellSciences), 05-172 (Upstate) and MAB291 (R&D Systems) showed different abilities toinhibit IGF binding to 1GFBP with IC50 values ranging from low ng/mL to 100 ng/mL. Noinhibition of IGF-I binding to the IGFBP-3 was observed with irrelevant mouse IgG andhuman IgG up to 10 pg/mL, suggesting that the anti-IGF-I effect is specific. Commerciallyavailable monoclonal antibodies 05-166 (Upstate) and MAB292 (R&D) showed a significantdifference in affinity for inhibition of IGF-II / IGFBP-3 interactions. These experimentsshow that anti-IGF mAbs can block the binding of IGF to IGFBP-3, giving an assay thatcould be used for screening purified antibodies from hybridoma lines.. The next step was toevaluate the effects of exhausted hybridoma medium on the assay signal.

[0177] Serial dilutions of the hybridoma medium and anti-KLH hybridomaexhaust supernatants were tested in the assay system. When hybridoma supernatants werediluted 1:10 in preparation for preincubation with 1GFI/II (final dilution in the assay was 53 WO 2007/070432 PCT/US2006/047059 1:60), there was almost no effect of the medium on the assay results. Based on these data, hybridoma supernatants were diluted for preincubation with IGF, providing the preferred 1/60 dilution final dilution in the assay.

[0178] Six hundred eighty-three exhaust supernatants positive for IGF-1 and IGF-II binding were examined for their ability to inhibit binding of IGF to 1GFBP-3. Inhibitionabove 50% for IGF-I and above 60% for IGF-II were used as cut-off criteria. The summaryresults of the screen using these cut-offs are shown in Table 6.

TABLE 6. NUMBERS OF POSITIVE HITS IDENTIFIED IN THE SCREEN IGF-I IGF-II 1GF-1/11 Samples Inhibitions >50% >60% 376 (plates 1-4) 48 51 19 307 (plates 5-8) 39 78 32 683 Total 87 129 51 [0179] The IGFBP competition assay using the AlphaScreen assay identified 87samples inhibiting IGF-I binding to IGFBP-3 and 129 samples inhibiting IGF-II binding toIGFBP-3 among 683 tested supernatants. Fifty-one samples demonstrated dual competitionof IGF-I and IGF-II. However, in order to more carefully reproduce the function or behaviorof the antibodies in vivo, where the IGF and the IGFBP complex would be largely preformed,additional assays, as described in example 8 were performed. EXAMPLE 7

DETERMINATION OF ANTI-IGF-1 AND IGF-II ANTIBODY AFFINITY USING BIACORE ANALYSIS (LOW RESOLUTION SCREEN)

Low Resolution Screen of 34 Purified Monoclonal Antibodies [0180] The label-free surface piasmon resonance (SPR), or Biacore, was utilizedto measure the antibody affinity to the antigen. For this purpose, a high-density goat anti-human antibody surface over a CM5 Biacore chip was prepared using routine aminecoupling. All the mAbs were diluted to approximately 20 pg/ml in HBS-P running buffercontaining 100 pg/ml BSA. Each mAb was captured on a separate surface using a 30-secondcontact time at 10 pL/min., and a 5-minute wash for stabilization of the mAb baseline.

[0181] IGF-I was injected at 335.3 nM over all surfaces at 23°C for 120 seconds,followed by a 5-minute dissociation, using a flow rate of 100 pL/min. The samples were 54 WO 2007/070432 PCT/US2006/047059 prepared in the HBS-P running buffer described above. The surfaces were regenerated afterevery capture/injection cycle with one 15-second pulse of 146mM phosphoric acid (pH 1.5).The same capture/injection cycles were repeated for each antibody with 114.7 nM IGF-II.Drift-corrected binding data for the 34 mAbs was prepared by subtracting the signal from acontrol flow cell and subtracting the baseline drift of a buffer injected just prior to eachantigen injection. Data were fit globally to a 1:1 interaction model using CLAMP todetermine the binding kinetics (David G. Myszka and Thomas Morton (1998) “CLAMP©: abiosensor kinetic data analysis program,” TIBS 23, 149-150). A mass transport coefficientwas used in fitting the data. The kinetic analysis results of IGF-1 and IGF-II binding at 25°Care listed in Table 7 below. The mAbs are ranked from highest to lowest affinity.

TABLE 7. IGF-1 AND IGF-II LOW RESOLUTION BIACORE SCREEN OF 34MONOCLONAL ANTIBODIES IGF-II IGF-I Sample ka (NT’s’1) kd (s’) Kn(pM) ka (M-'s-1) kd (s-1) Kn(pM) 7.159.2 3.5 X 106 1.0 X IO’5* 2.9 4.3 X IO6 9.3 X IO-4 216.0 8.86.1 6.1 X 106 2.4 X I0“4 39.3 3.4 X 106 1.3 X IO’2 3823.0 4.25.1 9.3 X 106 4.1 X IO'4 44.1 5.1 X 106 6.9 X 10'3 1353.0 7.234.2 6.4 X 106 2.9 X IO’4 45.3 6.7 X 106 2.2 X IO'3 328.0 7.160.2 4.6 X 106 2.5 X 10-4 54.3 5.6 X 106 3.3 X 10'3 589.0 7.146.3 3.2 X 106 1.8 X IO"4 56.2 3.8 X 106 8.7 X 10'3 2289.0 7.34.1 3.0 X 106 1.8 X 10'4 60.0 5.2 X 106 3.2 X 10‘3 615.0 7.123.1 4.0 X 106 3.4 X IO"4 85.0 4.3 X 106 9.0 X 10 3 2093.0 7.202.3 1.8 X 106 1.7 X IO’4 94.4 1.2 X 106 5.4 X 10’3 4500.0 4.141.1 4.9 X 106 4.7 X IO"4 95.9 * A A 7.215.2 3.2 X 106 3.3 X 10-4 103.0 3.6 X 106 2.6 X IO'2 7222.0 8.287.2 2.4 X 106 2.5 X IO"4 104.0 7.7 X 10s 1.3 X IO’3 1688.0 8.146.2 7.7 X 106 8.0 X 10’4 104.0 2.4 X 106 1.3 X IO’2 5417.0 4.143.2 1.1 X 107 1.2 X IO’3 109.0 6.4 X 106 1.9 X IO'2 2969.0 7.251.3 3.5 X 106 4.3 X IO'4 123.0 4.6 X 106 4.3 X 10'3 935.0 7.99.1 6.9 X 106 9.9 X 104 143.0 6.0 X 106 4.8 X 10*3 800.0 55 WO 2007/070432 PCT/US2006/047059 IGF-II IGF-I Sample k„ (MS ') kd (s'1) K„(pM) ka (M-’s-’) kd (s'’) KD(pM) 4.142.2 5.3 X 106 8.5 X 10"4 160.0 8.5 X 106 1.9 X IO'2 2235.0 7.41.3 3.2 X 106 5.5 X IO'4 172.0 5.2 X 106 2.9 X 103 558.0 7.56.3 3.3 X 106 6.0X W4 182.0 4.8 X 106 3.1 X IO'3 646.0 7.127.1 4.1 X 106 7.6 X 10·4 185.0 4.9 X IO6 3.5 X IO'3 714.0 8.8.3 4.0 X 106 7.8 X 10'4 195.0 3.1 X 106 8.2 X 10'4 264.0 7.158.2 3.4 X 106 6.7 X IO'4 197.0 4.4 X 106 2.2 X 10'3 500.0 7.23.3 3.3 X 106 6.5 X 1 O'4 197.0 4.1 X 106 5.8 X IO'3 1415.0 7.252.1 3.2 X 106 6.5 X IO'4 203.0 2.3 X 106 2.1 X IO'3 913.0 7.66.1 3.6 X 106 7.7 X 10'4 214.0 2.0 X 106 2.5 X IO'3 1250.0 7.130.1 4.2 X 106 1.0 X 10'3 238.0 4.6 X 106 1.8X 10'2 3913.0 4.90.2 4.9 X 106 1.2 X 10'3 245.0 * * * 7.3.3 3.2 X 106 8.8 X 10'4 275.0 4.1 X 106 3.9 X IO3 951.0 7.118.1 4.6 X 106 1.5 X 10'3 326.0 5.6 X 106 5.1 X IO'3 911.0 7.212.1 4.2 X 106 1.6 X 10'3 381.0 3.1 X 106 6.0 X IO'3 1935.0 7.175.2 1.3 X 106 7.4 X 10’4 569.0 1.8 X 106 2.0 X 10'2 11111.0 4.121.1 5.5 X 106* 3.2 X IO'3* 582.0* 1.5 X 106 2.1 X IO'3 1400.0 7.85.2 1.9 X 106 1.3 X 10'3 684.0 2.2 X 106 2.9 X IO'2 13182.0 7.58.3 8.9 X 106 7.9 X IO'3 888.0 7.2 X 106 3.2 X IO'2 4444.0 IGF-I Binding Data [0182] Most mAbs fit a 1:1 model reasonably well. MAbs 4.90.2 and 4.141.1were characterized by extremely complex data. These mAbs were listed with an asterisk inTable 7 because no meaningful kinetic constants could be estimated from the 1:1 model Fit.The latter off-rate phase appears to be very slow for both of these mAbs (at least 1 X 10'5sec'1), which might make these two mAbs useful as therapeutic compounds. 56 WO 2007/070432 PCT/US2006/047059 IGF-I1 Binding Data [0183] Most mAbs fit a 1:1 model reasonably well. The off-rate for mAb 7.159.2was held constant at 1 X 1 O'5 sec"1 because there was not enough decay data to adequatelyestimate ka.

[0184] The low-resolution Biacore studies in this example are designed as a semi-quantitative ranking approach. In order to acquire more accurate information regarding thecharacteristic rate constants and affinities of individual mAbs, high-resolution Biacorestudies were carried out as described in Example 8. EXAMPLE 8 DETERMINATION OF ANTI-IGF-I AND IGF-11 ANTIBODY AFFINITY USINGBIACORE ANALYSIS (HIGH RESOLUTION SCREEN) [0185] A high resolution Biacore analysis was performed to further measure theantibody affinity to the antigen. mAbs 7.159.2, 7.234.2, 7.34.1, 7.251.3, and 7.160.2 wereeach captured and the IGF-I and IGF-11 antigens were each injected over a range ofconcentrations. The resulting binding constants are listed in Table 8.

TABLE 8. ANTI-IGF ANTIBODY AFFINITY DETERMINED BY LOW-AND HIGH-RESOLUTION BIACORE ANALYSIS mAb Low resolution KD (pM) High Resolution KD (pM) IGF-I IGF-II IGF-I IGF-II 7.159.2 216.0 2.9 294.0 1.9 7.234.2 328.0 45.3 3760.0 295.0 7.34.1 615.0 60.0 436.0 421.0 164.0 162.0 7.251.3 935.0 123.0 452.0 47.4 7.160.2 589.0 54.3 2800.0 237.0 57 WO 2007/070432 PCT/US2006/047059 [0186] Thus, embodiments of the invention can include an antibody that willpreferentially bind to IGF-II, but that will cross-react with IGF-I, binding to IGF-II withhigher affinity than to IGF-I. For example, the antibody can bind to IGF-II with 2.5 timesgreater affinity than to IGF-I. In certain embodiments, the antibody can bind to IGF-II withat least 5, at least 10, at least 25, at least 50 or at least 150 times greater affinity than to IGF-I.

Screening of Preformed IGF-I/GFBP-3 Complexes [0187] The IGFBP competition assay described in Example 6 identified 87samples inhibiting IGF-I binding to IGFBP-3 and 129 samples inhibiting IGF-II binding toIGFBP-3 among 683 tested supernatants. Fifty-one samples demonstrated dual competitionof IGF-I and IGF-II. However, in order to more carefully reproduce the function or behaviorof the antibodies in vivo, where the IGF and the IGFBP complex would be largely preformed,the following Biacore assays were performed on selected antibodies.

[0188] Six selected antibodies were screened to determine whether they bindIGF-1 or IGF-II in complex with IGFBP. All six of the selected mAbs (7.159.2, 7.146.3,7.34.1, 7.251.3, 7.58.3, and unrelated control antibody ABX-MA1) were covalentlyimmobilized to a high surface capacity (5,400-12,800 RUs) on two CM5 Biacore chips usingroutine amine coupling with a Biacore 2000 instrument. One flow cell on each CM5 chipwas activated and blocked (no mAb immobilized) for use as a control surface.

[0189] Next, IGF-I and IGFBP-3 were mixed together in Hepes buffered saline,pH 7.4, 0.005% P-20, 100 pg/ml BSA (HBS-P), to make a final solution of 193 nM and 454nM, respectively. IGF-II and IGFBP-3 were mixed together to make a final solution of 192nM and 455 nM, respectively. Under these conditions, IGF-I and IGF-II were 99.97%complexed by IGFBP-3. Equilibrium was reached within minutes under these conditions.Solutions of complexed 1GF-I/IGFBP-3 and IGF-II/IGFBP-3 were flowed across the variousmAb surfaces at 40 pL/min and 23 °C, for 180 seconds and dissociation was followed for120 seconds. Uncomplexed IGF-I and IGF-II were then flowed across each surface at 193nM and 192 nM, respectively, and IGFBP-3 was flowed across each surface at 454 nM. Thesurfaces were regenerated with a 20 second pulse of 10 mM glycine, pH 2.0. 58 WO 2007/070432 PCT/US2006/047059 [0190] The sensorgrams were processed using the program Scrubber bysubtracting the bulk refractive index change and any nonspecific binding signal of the analyteto the blank surface from the binding signal from surfaces with mAb immobilized. Afterblank correction subtraction, the sensorgrams were referenced a second time by subtractingan average sensorgram for buffer injections over a specific flow cell. This “doublereference” corrected the mAb binding sensorgrams for any systematic errors present on aparticular flow cell.

[0191] Complexed and uncomplexed IGF-I/IGFBP-3 and 1GF-II/IGFBP-3 boundfairly weakly to the bound antibodies, with a rough estimate of the nonspecific bindinginteraction being a Kd>1 μΜ for all six mAbs, including negative control ABX-MA1 (SeeTable 9). However, with ABX-MA1 the IGF-I/II binding was weak and indicatednonspecific binding interactions occurred with all these three analytes. Apparently, theIGF/IGFBP-3 complexes bind slightly stronger to all these mAbs than IGFBP-3 does alone.However, because both IGF-I, IGF-II and IGBP-3 appear to bind nonspecifically to thesemAbs themselves, when they are both bound together, this results in an even “stickier”nonspecific binding protein complex, which explains the greater binding signal for thecomplex. The IGF-I/II/IGFBP-3 complexes and IGFBP-3 bound to the control surfacesignificantly also indicating the nonspecificity of these two proteins. However, in thesensorgrams below this background binding is subtracted out in the first reference duringdata processing, as described above.

[0192] This experiment suggests that although 51 of the samples were previouslyshown to inhibit binding of IGF-I/II to IGFBP3 (Example 6), the antibodies may also bind tothe IGF/1GFBP complex in vitro. TABLE 9. BINDING SUMMARY FOR IGF-I/IGFBP-3 AND IGF-II/IGFBP-3 BINDINGTO SIX MABS. 59 WO 2007/070432 PCT/US2006/047059 inAb IGF-I/IGFBP-3 complex IGF-II/IGFBP-3 complex IGFBP-3 IGF-I (or II) 7.159.2 4* + 4“ +++ 7.146.3 + ++ 4- +++ 7.34.1 + ++ + +++ 7.251.3 ++ ++ ++ +++ 7.58.3 ++ ++ ++ 4-4-4- ABX-MA1 + + + + +, slight binding relative to IGF-I or IGF-II to the mAb++, medium binding relative to IGF-I or IGF-II to the mAb+++, strong binding relative to IGF-I or IGF-II binding to the mAb♦These ratings DO NOT indicate the Kd for these interactions. EXAMPLE 9

DETERMINATION OF ANTI-INSULIN ANTIBODY AFFINITY USING BIACORE ANALYSIS (LOW RESOLUTION SCREEN) [0193] The cross-reactivity of antibodies to IGF-I/II was further investigated bymeasuring the affinity of the mAbs to human insulin. IGF-I/II antibodies were immobilizedto the CM5 Biacore chips, and insulin in solution was injected for the determination of theon-rate and off-rate. Five mAbs, including 7.234.2, 7.34.1, 7.159.2, 7.160.2, and 7.251.3,were tested in this experiment. Insulin diluted to 502 nM in the running buffer was injectedover all capture surfaces.

[0194] No insulin binding to any of the mAbs was observed at 502 nM insulin.These results suggest that there is no apparent cross-reactivity of the IGF-I/II mAbs withinsulin. EXAMPLE 10

BINNING OF ANTIBODIES

[0195] Epitope binning was performed to determine which of the anti-IGF-I/11antibodies would cross compete with one another, and thus were likely to bind to the sameepitope on IGF-I/II. The binning process is described in U.S. Patent Application 60 191718/2 20030175760, also described in Jia et al., J. Immunol. Methods, (2004) 288:91-98. Briefly,Luminex beads were coupled with mouse anti-huIgG (Pharmingen #555784) following theprotein coupling protocol provided on the Luminex website. Pre-coupled beads wereprepared for coupling to primary unknown antibody using the following procedure,protecting the beads from light. Individual tubes were used for each unknown supernatant.The volume of supernatant needed was calculated using the following formula: (nX2+10)x 50 μΐ (where n = total number of samples). A concentration of 0.1 pg/ml was used in thisassay. The bead stock was gently vortexed, and diluted in supernatant to a concentration of2500 of each bead in 50 μΐ per well or 0.5X105 beads/ml.

[0196] Samples were incubated on a shaker in the dark at room temperatureovernight.

[0197] The filter plate was pre-wetted by adding 200 μΐ wash buffer per well,which was then aspirated. 50 μΐ of each bead was added to each well of the filter plate.Samples were washed once by adding 100 μΙ/well wash buffer and aspirating. Antigen andcontrols were added to the filter plate at 50 μΙ/well. The plate was covered, incubated in thedark for 1 hour on a shaker, and then samples were washed 3 times. A secondary unknownantibody was then added at 50 μΙ/well. A concentration of 0.1 μg/ml was used for theprimary antibody. The plate was then incubated in the dark for 2 hours at roomtemperature on a shaker, and then samples were washed 3 times. 50 μΙ/well of biotinylatedmouse anti-human IgG (Pharmingen #555785) diluted at 1:500 was added, and sampleswere incubated in the dark for 1 hour with shaking at room temperature.

[0198] Samples were washed 3 times. 50 μΙ/well Streptavidin-PE at a 1:1000dilution was added, and samples were incubated in the dark for 15 minutes with shaking atroom temperature. After running two wash cycles on the LuminexlOO, samples werewashed 3 times. Contents in each well were resuspended in 80 μΐ blocking buffer. Sampleswere carefully mixed with pipetting several times to resuspend the beads. Samples werethen analyzed on the LuminexlOO. Results are presented below in Table 10. 61

I ! P/15804/125647/596826/l WO 2007/070432 PCT/US2006/047059

TABLE 10. BINS FOR TOP 34 IGF-I/II ANTIBODIES POSITIVE IN FUNCTIONAL

ASSAY IGF-I Bin 1 Bin 2 Bin 3 No Bin 7.3.3 7.58.3 7.175.2 7.215.2 7.23.3 8.287.2 7.85.2 7.66.1 4.90.2 7.56.3 4.141.1 7.160.2 7.146.3 7.41.3 7.34.1 4.121.1 7.159.2 8.146.2 7.251.3 7.252.1 7.123.1 7.212.1 7.234.2 7.99.1 7.127.1 4.25.1 8.8.3 7.158.2 7.202.3 7.130.1 8.86.1 4.142.2 7.118.1 4.143.2 IGF-II Bin 1 Bin 2 Bin 3 No Bin 7.3.3 7.158.2 7.175.2 7.215.2 7.127.1 8.146.2 4.90.2 7.99.1 7.252.1 4.141.1 7.123.1 8.86.1 7.85.2 7.212.1 7.251.3 7.234.2 7.159.2 7.130.1 7.146.3 7.118.1 7.34.1 8.287.2 7.58.3 • 7.66.1 7.41.3 7.56.3 7.160.2 7.202.3 8.8.3 4.25.1 7.23.3 4.142.2 4.143.2 4.121.1 EXAMPLE 11

STRUCTURAL ANALYSIS OF ANTI-1GF-I/H ANTIBODIES

[0199] The variable heavy chains and the variable light chains of severalantibodies were sequenced to determine their DNA sequences. The complete sequenceinformation for the anti-IGF-I/II antibodies is provided in the sequence listing withnucleotide and amino acid sequences for each gamma and kappa chain, combination. Thevariable heavy sequences were analyzed to determine the VH family, the D-region sequenceand the J-region sequence. The sequences were then translated to determine the primary 62 WO 2007/070432 PCT/US2006/047059 amino acid sequence and compared to the germline VH, D and J-region sequences to assess somatic hypermutations.

[0200] The alignment of the sequences of these antibodies to their germline genesare shown in the foilowing tables. Table 11 is a table comparing the antibody heavy chainregions to their cognate germ line heavy chain region. Table 12 is a table comparing theantibody kappa light chain regions to their cognate germ line light chain region. Mutationsaway from germline are shown as the new amino acid.

[0201] The variable (V) regions of immunoglobulin chains are encoded bymultiple germ line DNA segments, which are joined into functional variable regions (VhDJhor VkJk) during B-cell ontogeny. The molecular and genetic diversity of the antibodyresponse to IGF-I/II was studied in detail. These assays revealed several points specific toanti-IGF-I/II antibodies.

[0202] Analysis of five individual antibodies specific to IGF-I/II resulted in thedetermination that the antibodies were derived from three different germline VH genes, fourof them from the VH4 family, with 2 antibodies being derived from the VH4-39 genesegment. Tables 11 and 12 show the results of this analysis.

[0203] It should be appreciated that amino acid sequences among the sister clonescollected from each hybridoma are identical. For example, the heavy chain and light chainsequences for mAb 7.159.2 are identical to the sequences shown in Tables 11 and 12 formAb 7.159.1.

[0204] The heavy chain CDRls of the antibodies of the invention have asequence as disclosed in Table 11. The CDRls disclosed in Table 11 are of the Kabatdefinition. Alternatively, the CDRls can be defined using an alternative definition so as toinclude the last five residues of the FR1 sequence. For example, for antibody 7.159.1 theFR1 sequence is QVQLVQSGAEVKKPGASVKVSCKAS (SEQ ID NO.: 93) and the CDR1sequence is GYTFTSYDIN (SEQ ID NO.: 94); for antibody 7.158.1 the FR1 sequence isQLQLQESGPGLVKPSETLSLTCTVS (SEQ ID NO.: 95) and the CDR1 sequence isGGS1RSSSYYWG (SEQ ID NO.: 96); for antibody 7.234.1 the FR1 sequence isQLQLQESGPGLVKPSETLSLTCTVS (SEQ ID NO.: 97) and the CDR1 sequence isGGSINSSSNYWG (SEQ ID NO.: 98); for antibody 7.34.1 the FR1 sequence isQVQLQESGPGLVKPSETLSLTCTVS (SEQ ID NO.: 99) and the CDR1 sequence is 63 WO 2007/070432 PCT/US2006/047059 GGSISSYYWS (SEQ ID NO.: 100); and for antibody 7.251.3 the FRI sequence is QVQLQESGPGLVKPSETLSLTCTVS (SEQ ID NO.: 101) and the CDR1 sequence is GGSISSYYWS (SEQ ID NO.: 102).

[0205] It should also be appreciated that where a particular antibody differs fromits respective germline sequence at the amino acid level, the antibody sequence can bemutated back to the germline sequence. Such corrective mutations can occur at one, two,three or more positions, or a combination of any of the mutated positions, using standardmolecular biological techniques. By way of non-limiting example, Table 12 shows that thelight chain sequence of mAb 7.34.1 (SEQ ID NO.: 12) differs from the correspondinggermline sequence (SEQ ID NO.:80) through a Pro to Ala mutation (mutation 1) in the FRIregion, and via a Phe to Leu mutation (mutation 2) in the FR2 region. Thus, the amino acidor nucleotide sequence encoding the light chain of mAb 7.34.1 can be modified to changemutation 1 to yield the germline sequence at the site of mutation 1. Further, the amino acidor nucleotide sequence encoding the light chain of mAb 7.34.1 can be modified to changemutation 2 to yield the germline sequence at the site of mutation 2. Still further, the aminoacid or nucleotide sequence encoding the light chain of mAb 7.34.1 can be modified tochange both mutation 1 and mutation 2 to yield the germline sequence at the sites of bothmutations 1 and 2. 64

TABLE 11. HEAVY CHAIN ANALYSIS

Chain Name SEQ ID NO. V D J FR1 CDR1 FR2 CDR2 FR3 CDR3 FR4 Germ- line** 75 VH1-8 N.A. JH6B QVQLVQSGAEVKKPG ASVKVSCKASGYTFT SYDI N WVRQATG QGLEWMG WMNPNSGNT GYAQKFQG RVTMTRNTSISTAYMELS SLRSEDTAVYYCAR ##YYYYY GMDV WGQGTT VTVSSA 7_159_1 6 tt tt tt QVQLVQSGAEVKKPG ASVKVSCKASGYTFT SYDI N WVRQATG QGLEWMG WMNPNSGNT GYAQKFQG RVTMTRNTSISTAYMELS SLRSEDTAVYYCAR DPYYYYY GMDV WGQGTT VTVSSA Germline 77 VH4-39 D6- 19 JH2 QLQLQESGPGLVKPS ETLSLTCTVSGGSIS SSSY YWG WIRQPPG KGLEWIG SIYYSGSTY YNPSLKS RVTISVDTSKNQFSLKLS SVTAADTAVYYCAR ####ss# #WYFDL WGRGTL VTVSSA 7_158_1 2 tt tt tt QLQLQESGPGLVKPS ETLSLTCTVSGGSIR SSSY YWG WIRQPPG KGLEWIG GIYYSGSTY YNPSLKS RVTMSVDTSKNQFSLKLS SVTAADTAVYYCAR QRGHSSG WWYFDL WGRGTL VTVSSA 7_234_1 18 tt tt tt QLQLQESGPGLVKPS ETLSLTCTVSGGSIN SSSN YWG WIRQPPG KGLAWIG GIYYSGSTY YNPSLRS RVTMSVDTSKNQFSLKLS SVTAADTAVYYCAR QRGHSSG WWYFDL WGRGTL VTVSSA Germline 79 VH4-59 Dl- 20 JH6B QVQLQESGPGLVKPS ETLSLTCTVSGGSIS SYYW S WIRQPPG KGLEWIG YIYYSGSTN YNPSLKS RVTISVDTSKNQFSLKLS SVTAADTAVYYCA#R ITGT### GMDV WGQGTT VTVSSA 7_34_1 10 tt tt tt QVQLQESGPGLVKPS ETLSLTCTVSGGSIS SYYW s WIRQPPG RGLEWIG YFFYSGYTN YNPSLKS RVTMSVDTSKNQFSLKLS SVTAADTAVYYCAC ITGTTKG GMDV WGQGAT VTVSSA 7_251_3 14 tt tt tt QVQLQESGPGLVKPS ETLSLTCTVSGGSIS SYYW S WIRQPPG KGLEWIG YFFYSGYTN YNPSLKS RVTISVDTSKNQFSLKLS SVTAADTAVYYCAC ITGTTKG GMDV WGQGTT VTVSSA * The hatch designation (#) indicates a space in the germline and is used to show a proper alignment with the antibody sequences shown in the table. ** The germline sequences shown in the above table are for alignment purposes, and it should be realized that each individualantibody region exists in its own location within the variable regions of immunoglobulin germline DNA segments in vivo. WO 2007/070432 PCT/US2006/047059

TABLE 12. LIGHT CHAIN ANALYSIS

Chain Name SEQ ID NO. V J FR1 CDR1 FR2 CDR2 FR3 CDR3 FR4 Germlin e 76 Vl-19 JL 2 QSVLTQPPSVSA APGQKVTISC SGSSSNI GNNYVS WYQQLPGT APKLLIY DNNKRPS GIPDRFSGSKSGTSAT LGITGLQTGDEADYYC GTWDSSL SA##V FGGGTK LTVLG 7__159_1 8 n II QSVLTQPPSVSA APGQKVTISC SGSSSNI ENNHVS WYQQLPGT APKLLIY DNNKRPS GIPDRFSGSKSGTSAT LGITGLQTGDEADYYC ETWDTSL SAGRV FGGGTK LTVLG Germlin e 78 L5 JK 3 DIQMTQSPSSVS ASVGDRVTITC RASQGIS SWLA WYQQKPGK APKLLIY AASSLQS GVPSRFSGSGSGTDFT LTISSLQPEDFATYYC QQANSFP FT FGPGTK VDIKR 7_158_1 4 tt «1 DIQMTQSPSSVS ASVGDSVTITC RASQGIS SYLA WYQQKPGK APKLLIY AASSLQS GVPSRFSGNGSGTDFT LTISSLQPEDFATYYC QQANNFP FT FGPGTK VDIKR 7_234_1 20 11 It DIQMTQSPSSVS ASVGDRVTITC RASRGIS SWLA WYQQRPGK APKLLIY TASSLQS GVPSRFSGSGSGTDFT LTIS SLQPEDFATYYC QQANSFP FT FGPGTK VDIKR Germline 80 Vl-13 JL 2. QSVLTQPPSVSG APGQRVTISC TGSSSNI GAGYDVH WYQQLPGT APKLLIY GNSNRPS GVPDRFSGSKSGTSAS LAITGLQAEDEADYYC QSYDSSL SGSV FGGGTK LTVLG 7_34_1 12 n II QSVLTQAPSVSG APGQRVTISC TGRSSNI GAGYDVH WYQQFPGT APKLLIY GNSNRPS GVPDRFSGSKSGTSAS LAITGLQAEDEADYYC QSYDSSL SGSV FGGGTK LTVLG 7_251_3 16 n tl QSVLTQPPSVSG APGQRVTISC TGSSSNI GAGYDVH WYQQLPGT APKLLIY GNNNRPS GVPDRFSGSKSGTSAS LAITGLQADDEADYYC QSFDSSL SGSV FGGGTK LTVLG * The hatch designation (#) indicates a space in the germline and is used to show a proper alignment with the antibody sequences shown in the table. ** The germline sequences shown in the above table are for alignment purposes, and it should be realized that each individualantibody region exists in its own location within the variable regions of immunoglobulin germline DNA segments in vivo. WO 2007/070432 PCT/US2006/047059 WO 2007/070432 PCT/US2006/047059 EXAMPLE 12

INHIBITION OF IGF-I AND IGF-II-INDUCED PHOSPHORYLATION OF hIGF-IR

ECTOPICALLY EXPRESSED IN NIH3T3 CELLS

[0206] IGF ligands exert their proliferation and anti-apoptosis functions byactivating receptor tyrosine kinase activity in the IGF-IR receptor. In order to evaluatethe anti IGF-I/II antibodies for their ability to inhibit IGF-induced phosphorylation ofIGF-IR, NIH3T3 cells ectopically expressing hIGF-IR, were used in the following assay.

[0207] NIH3T3 cells ectopically expressing the human IGF-IR were seeded ina 96-well plate at a density of 10,000 cells per well and incubated overnight in starvationmedia (1% charcoal stripped FBS). The following day, the growth medium wasdiscarded, the wells were gently washed twice with PBS, and lOOpL of serum-freemedium (0% FBS) was added to starve the cells. After 1-2 hours, lOOul of serum-freemedium with 0.05% BSA containing either IGF-I (lOnM) or IGF-II (lOnM) that was pre-incubated for 60 minutes at 37°C with various antibody concentrations, was added to thecells in triplicate. The stimulation was allowed to occur for 10 minutes at 37°C, afterstimulation, media removed and lOOuL 3.7%fromaldehyde in PBS/3%BSA added to eachwell and incubated at RT for 20 min. The cells were then washed 2X with PBS andlOOuL permeabilization buffer (0.1% Triton-X in 3%BSA/PBS) was added to each well.This was allowed to incubate at RT for 10 min, discarded and lOOul of 0.6% hydrogenperoxide in PBS/3% BSA was added to inactivate any endogenous peroxidase activity.After a 20min RT incubation, the cells were washed 3X with PBS/0.1% Tween-20 andblocked by adding lOOuL 10%FBS in PBS/0.1% Tween-20 at RT for lhr. The BlockingBuffer was then removed and 50uL anti-phospho IGFIR antibody at lug/ml (cat#44-804,BioSource) was added to each well in 10%FBS/PBS-T. After a 2hr RT incubation cellswere washed 3X with PBST soaking for 5 minutes between each wash. After the washes50ul/well of a Goat anti Rabbit IgGFc-HRP secondary antibody diluted 1:250 in BlockingBuffer was added to each of the well. After a 1 hour RT incubation the cells were washed3X for 5 minutes with PBST as before and tapped dry. 50ul of ECL reagent (DuoLux)was then added and RLUs was read immediately. 67 WO 2007/070432 PCT/US2006/047059 [0208] Thirty-two (32) antibody lines were screened, and two independentassays were performed for each antigen. The results for the top ten antibodies aresummarized in Table 13 below.

TABLE 13. SUMMARY OF INHIBITION OF IGF-DEPENDENT IGF-IRPHOSPHORYLATION IN NIH3T3 CELLS IGF-I pTYR Results (n=2) IGF-II pTYR Results (n=2) % MAXActivation EC50 (nM)* % MAXActivation EC50 (nM)* mAb ID Mean SD Mean SD Mean SD Mean SD 7.159.2 16.5% 6.6% 7.4 0.8 28.6% 5.4% 3.1 0.2 7.34.1 14.2% 1.8% 9.4 0.8 21.5% 3.9% 2.5 0.1 7.146.3 19.5% 3.9% 19.0 5.7 23.6% 2.1% 3.6 0.2 7.251.3 16.9% 5.4% 14.5 0.7 15.9% 1.5% 3.0 0.9 7.234.2 21.1% 3.0% 24.3 1.0 21.1% 0.1% 7.7 0.1 7.160.2 33.6% 6.4% 22.9 0.1 21.5% 0.6% 4.7 0.2 7.158.2 22.7% 0.9% 28.3 0.5 33.7% 2.4% 11.3 3.2 7.56.3 31.3% 2.2% 25.1 4.3 21.2% 0.2% 6.3 0.5 7.118.1 24.1% 5.2% 40.8 2.6 21.8% 3.2% 13.9 5.3 7.41.3 33.1% 6.1% 47.1 7.0 29.5% 4.4% 3.5 4.0

* These assays may have been run under antigen limiting conditions given the mAb KD for IGF-I and IGF-II. EXAMPLE 13

INHIBITION OF IGF-I AND IGF-II-INDUCED PROLIFERATION OF NIH3T3CELLS TRANSFECTED WITH HIGF-IR

[0209] As discussed above, one of the criteria for neutralizing IGF-I/IIantibodies is the ability to inhibit IGF-induced proliferation. In order to evaluate theantibodies for their ability to inhibit IGF-induced proliferation, NIH3T3 cells ectopicallyexpressing hIGF-IR, were used in the following assay.

[0210] NIH3T3 cells ectopically expressing hIGF-IR were seeded in a 96-wellplate at a density of 5000 cells per well and cultured overnight in starvation medium (1%charcoal stripped FBS). The following day, the growth medium was discarded, the wellswere gently washed twice in medium without serum, and 100pL of serum-free mediumwas added to starve the cells. 100pL of starvation media containing 15ng/ml IGFI or50ng/ml IGF1I pre-incubated for 30 min at 37°C with various antibody concentrationswas added to the cells in duplicate or triplicate. Following a 20hr incubation cells are 68 WO 2007/070432 PCT/US2006/047059 pulsed with BrdU for 2hrs and the degree of incorporation (proliferation) was quantitatedusing the Cell Proliferation ELISA kit from Roche (Roche, Cat#l 647 229).

[0211] A total of 32 antibody lines were screened, and two or threeindependent assays were performed for each antigen. The results for the top 10antibodies are summarized in Table 14 below.

TABLE 14. SUMMARY OF INHIBITION OF IGF-DEPENDENT PROLIFERATIONOF NIH3T3/hIGF-IR CELLS IGF-I Proliferation Assay IGF-II Proliferation Assay % Inhibition EC50 (nM)* % Inhibition EC50 (nM)* mAb ID Mean (n=3) SD Mean (n=3) SD Mean (n=2) SD Mean (n=2) SD 7.159.2 77.0% 9.6% 24.1 5.9 99.3% 0.6% 7.6 2.5 7.34.1 72.6% 5.6% 23.4 8.1 73.6% 11.8% 16.3 0.4 7.146.3 65.3% 5.5% 37.2 4.5 82.0% 6.9% 15.9 3.4 7.251.3 72.4% 15.3% 38.9 4.3 79.2% 7.8% 22.0 3.7 7.234.2 67.3% 6.9% 40.6 4.6 62.1% 17.2% 24.3 2.4 7.160.2 62.8% 5.7% 47.6 10.7 45.9% 0.8% 24.7 2.8 7.158.2 57.4% 19.5% 42.8 1.7 54.6% 6.6% 36.0 4.2 7.56.3 50.2% 7.8% 65.7 31.9 48.0% 10.9% 38.3 7.5 7.118.1 59.4% 14.5% 1626.6 2714.5 68.3% 0.8% 49.9 3.8 7.41.3 29.5% 14.7% 76.3 35.9 51.9% 13.7% 61.9 23.7

♦These assays may have been run under antigen limiting conditions given the mAb KD for IGF-I and IGF-II. EXAMPLE 14

INHIBITION OF IGF-I AND IGF-II-INDUCED PHOSPHORYLATION OF hIGF-IR

EXPRESSED IN BxPC3 HUMAN PANCREATIC TUMOR CELLS

[0212] IGF-I/II exert their proliferation and anti-apoptosis functions byactivating receptor tyrosine kinase activity in the IGF-IR receptor. In order to evaluatethe antibodies for their ability to inhibit IGF-induced phosphorylation of IGF-IR, BxPC3human pancreatic tumor cells, which express endogenous hIGF-IR, were used in thefollowing assay.

[0213] BxPC3 cells were seeded in a 96-well plate at a density of 55,000 cellsper well and incubated overnight in regular growth medium. The following day, thegrowth medium was discarded, the wells were gently washed twice in medium withoutserum, and 100pL of serum-free medium was added to starve the cells. After 24 hours, 69 WO 2007/070432 PCT/US2006/047059 the medium was discarded, and the cells were gently washed once in medium withoutserum. Serum-free medium with 0.05% BSA containing either IGF-I (20ng/ml) or IGF-II(75ng/ml) was pre-incubated for 30 minutes at 37°C with various antibodyconcentrations, and lOOpL was then added to the cells in triplicate. The plates wereincubated for 15 minutes at 37°C, and were subsequently rinsed with cold PBS. 1 OOpL oflysis buffer was added to the wells and the plates were incubated for 30 minutes at 4°C.The lysates were spun down at 2000 rpm for 10 minutes at 4°C, and the supernatant wascollected. IGF-IR phosphorylation was quantitated using the Duoset human phosphor-IGF-IR ELISA kit (R&D Systems, Cat. No. DYC1770).

[0214] Ten antibody lines were screened, and two independent assays wereperformed for each antigen. The results are summarized in Table 15 below.

TABLE 15. SUMMARY OF INHIBITION OF IGF-DEPENDENT IGF-IRPHOSPHORYLATION IGF-I pTYR Results (n=2) IGF-II pTYR Results (n=2) n=l n=2 n=l n=2 mAb ID Max %Inhibition(333.3nM) EC50 (nM) Max %Inhibition(333.3nM) EC50 (nM) Max %Inhibition(333.3nM) EC50 (nM) *Max % Inhibition (133.3nM) EC50 (nM) 7.159.2 100.0 3.3 100.0 1.6 100.0 1.6 91.2 1.7 7.34.1 100.0 5.9 98.5 3.8 100.0 2.0 89.7 1.9 7.146.3 96.4 16.1 94.2 10.7 100.0 2.0 87.9 2.0 7.251.3 95.7 7.5 95.2 5.3 100.0 N.D. 91.3 2.6 7.234.2 97.3 5.1 91.5 2.9 98.5 1.9 77.3 2.3 7.160.2 93.4 5.3 89.2 3.1 88.6 1.7 73.2 2.5 7.158.2 92.9 4.5 89.4 3.6 92.4 N.D. 74.0 4.2 7.56.3 84.9 N.D. 88.7 6.5 91.4 10.1 66.2 5.1 7.118.1 90.5 13.1 90.6 11.8 95.7 17.9 78.0 13.1 7.41.3 88.6 6.5 86.5 6.5 88.6 4.5 70.6 3.1 *333nM for the last 3 antibodies. N.D.: Not Determined EXAMPLE 15

INHIBITION OF IGF-I AND IGF-II-INDUCED PROLIFERATION OF BxPC3 HUMAN

PANCREATIC TUMOR CELLS

[0215] As discussed above, one of the criteria for neutralizing IGF antibodiesis the ability to inhibit IGF-induced proliferation. In order to evaluate the antibodies for 70 WO 2007/070432 PCT/US2006/047059 their ability to inhibit IGF-induced proliferation, BxPC3 human pancreatic tumor cells,which express endogenous hIGF-IR, were used in the following assay.

[0216] BxPC3 cells were seeded in a 96-well plate at a density of 2000 cellsper well and cultured overnight in regular growth medium. The following day, the growthmedium was discarded, the wells were gently washed twice in medium without serum,and lOOpL of serum-free medium with lOpg/ml transferrin and 0.1% BSA (assaymedium) was added to starve the cells. After 24 hours, the medium was discarded, thecells were gently washed once in medium without serum, and lOOpL of assay mediumcontaining 20ng/ml IGF preincubated for 30 min at 37°C with various antibodyconcentrations was added to the cells in duplicate or triplicate. The plates were incubatedfor 3 days, and proliferation was quantitated using the CellTiter-Glo reagent (Promega).

[0217] Ten antibody lines were screened, and two or three independent assayswere performed for each antigen. The results are summarized in Table 16 below. Basedon the functional data below and the data from the Example 14, the four best antibodieswere selected. IGF-I-induced proliferation assay data was excluded from the selectioncriteria because of the high assay variability observed.

TABLE 16. SUMMARY OF INHIBITION OF IGF-DEPENDENT PROLIFERATIONOF BxPC3 HUMAN PANCREATIC TUMOR CELLS IGF-II Proliferation Results (n=2) n=l n=2 MAb ID Max %Inhibition(333.3nM) EC50 (nM) *Max % Inhibition (133.3nM) EC50 (nM) 7.159.2 120.0 0.8 118.3 0.9 7.34.1 117.0 0.5 109.0 6.7 7.146.3 128.7 3.0 119.5 7.3 7.251.3 128.5 0.5 105.3 4.4 7.234.2 111.7 N.D. 200.7 2.6 7.160.2 79.7 1.1 155.7 N.D. 7.158.2 86.3 0.0013 148.3 N.D. 7.56.3 87.0 N.D. 112.3 102.0 7.118.1 114.0 34.0 137.0 54.7 7.41.3 102.0 N.D. 73.0 N.D. *333nM for the ast 3 antibodies N.D.: Not Determined 71 WO 2007/070432 PCT/US2006/047059 EXAMPLE 16

DETERMINATION OF CROSS-REACTIVITY WITH MOUSE IGF-I, IGF-II AND

INSULIN

[0218] One objective was to develop antibodies that were specific to IGF-Iand IGF-II but that have no cross-reactivity with insulin. In order to perform laterexperiments in animals, the antibodies should also cross-react with murine IGF-I/II butnot murine insulin. Accordingly, ELISA assays were performed to determine whetherselected antibodies were able to cross-react with murine IGFs or insulin. (0219] As shown in Table 17, five of the top ten antibodies were tested forcross-reactivity with mouse or rat insulin by ELISA. The ELISAs showed that theseantibodies had no cross-reactivity with mouse or rat insulin, compared to negative controlantibody PK16.3.1 and in contrast to positive control anti-rat insulin antibody.

TABLE 17. CROSS-REACTIVITY WITH MOUSE INSULIN OD 450 with different Ag Antibodies mouse Insulin Rat Insulin No Ag 7.159.2 0.52 0.52 0.56 7.160.2 0.60 0.57 0.62 7.34.1 0.48 0.47 0.55 7.251.3 0.55 0.53 0.56 7.234.2 0.51 0.49 0.66 Serum • 1.28 1.23 1.34 anti Rat Insulin 2.52 3.06 0.10 PK16.3.1 0.58 0.58 0.62 EXAMPLE 17

INHIBITION OF MOUSE IGF-I AND IGF-II-INDUCED PHOSPHORYLATION OF

HUMAN IGF-IR ECTOPICALLY EXPRESSED IN NIH3T3 CELLS

[0220] The monoclonal antibodies with cross-reactivity with mouse IGF-I andIGF-II were further tested in order to determine the extent they inhibit IGF-inducedphosphorylation of the IGF-IR. This assay was performed as previously described usingNIH3T3 cells ectopically expressing the hIGF-IR receptor. The results of this assay aresummarized in Table 18.

[0221] NIH3T3 cells ectopically expressing the human IGF-IR were seeded ina 96-well plate at a density of 10,000 cells per well and incubated overnight in starvationmedia (1% charcoal stripped FBS). The following day, the growth medium was 72 WO 2007/070432 PCT/US2006/047059 discarded, the wells were gently washed twice with PBS, and lOOpL of serum-freemedium (0% FBS) was added to starve the cells. After 1-2 hours, lOOul of serum-freemedium with 0.05% BSA containing either mouse IGF-I (lOnM) or IGF-Π (lOnM) (R&DSystems, Inc., Minneapolis, MN Cat. No. 791-MG and 792-MG respectively) that waspre-incubated for 60 minutes at 37°C with various antibody concentrations, was added tothe cells in triplicate. The stimulation was allowed to occur for 10 minutes at 37°C, afterstimulation, media removed and lOOuL 3.7% formaldehyde in PBS/3%BSA added toeach well and incubated at RT for 20 min. The cells were then washed 2X with PBS andlOOuL permeabilization buffer (0.1% Triton-X in 3%BSA/PBS) was added to each well.This was allowed to incubate at RT for 10 min, discarded and lOOul of 0.6% hydrogenperoxide in PBS/3% BSA was added to inactivate any endogenous peroxidase activity.After a 20min RT incubation, the cells were washed 3X with PBS/0.1% Tween-20 andblocked by adding lOOuL 10%FBS in PBS/0.1% Tween-20 at RT for lhr. The BlockingBuffer was then removed and 50uL anti-phospho IGFIR antibody at lug/ml (cat#44-804,BioSource) was added to each well in 10%FBS/PBS-T. After a 2hr RT incubation cellswere washed 3X with PBST soaking for 5 minutes between each wash. After the washes50ul/well of a Goat anti Rabbit IgGFc-HRP secondary antibody diluted 1:250 in BlockingBuffer was added to each of the well. After a 1 hour RT incubation the cells were washed3X for 5 minutes with PBST as before and tapped dry. 50ul of ECL reagent (DuoLux)was then added and RLUs was read immediately.

TABLE 18. INHIBITION OF MOUSE IGF-INDUCED PHOSPHORYLATION OFhIGF-IR

Mouse IGF-I EC50 (nM) Mouse IGF-II EC50 (nM) mAb ID n=l n=2 n=l n=2 7.159.2 2.8 5.7 3.1 5.0 7.34.1 6.0 10.2 4.0 9.7 7.251.3 6.7 10.6 5.4 8.7 7.234.2 46.0 36.1 7.160.2 49.5 225.2 EXAMPLE 18 73 WO 2007/070432 PCT/US2006/047059 INHIBITION OF THE GROWTH OF N1H3T3 CELLS EXPRESSING IGF-II AND IGF-

1R IN VIVO IN NUDE MICE

[0222] In order to evaluate the antibodies for their ability to inhibit IGF-II-induced proliferation in vivo, the following experiments were performed.

[0223] Female nude mice 6-8 weeks of age (supplied by Charles RiverLaboratories, Wilmington, MA, USA) were implanted subcutaneously with 5xl06 Clone32 cells (NIH3T3 cells ectopically overexpressing human IGF-II and human IGF-IR).The cells were suspended in PBS in a total inoculum volume of 330 μΐ. The tumors wereallowed to grow to 100-200mm3 prior to treatment with monoclonal antibodies 7.159.2,7.34.1 and 7.251.3. Antibodies or IgG2 isotype control antibody suspended in PBS wereadministered intraperitoneally to randomized groups of 9 or 12 mice weekly for 4 weeksat 5 or 50 mg/kg from Day 22. PBS was administered as a vehicle control to a furthergroup of 11 mice weekly for 4 weeks from Day 22. Tumor size and body weight wasmeasured 2-3 times per week. The results are summarized in Figures 1 and 2.

[0224] Significant tumor growth inhibition was observed (see Figure 1) withno significant weight loss occurring in any group (see Figure 2). Antibodies 7.159.2,7.34.1 and 7.251.3 significantly inhibited the growth of Clone 32 tumors at 5 and 50mg/kg/week (see Figure 1). EXAMPLE 19 INHIBITION OF THE GROWTH OF NIH3T3 CELLS EXPRESSING IGF-I AND IGF-

1R IN VIVO IN NUDE MICE

[0225] In the previous example, the antibodies were shown to inhibit 1GF-II-induced proliferation in vivo. In order to evaluate the antibodies for their ability to inhibitIGF-I-induced proliferation in vivo, the following experiments were performed.

[0226] Female nude mice (Alderley Park strain derived from Swiss nu/numice strain, supplied by AstraZeneca) were implanted with 5xl06 viable Pl2 cells[NIH3T3 cells ectopically overexpressing human IGF-I and human IGF-IR (Pietrzkowskiet al, Cell Growth & Differentiation, 3, 199-205, 1992)] subcutaneously in the left flank.The cells were suspended in PBS in a total inoculum volume of 0.1 ml. Two groups ofanimals (each n=10) were dosed twice weekly from day of NIH3T3 cell implant witheither mAb 7.159.2 at 1.0 mg per mouse or with an equivalent volume of PBS vehicle(0.3 ml) for the same schedule. All doses were given intraperitoneally via the (i.p.) route.Animal body weights were measured daily and, once established, tumor measurements 74 WO 2007/070432 PCT/US2006/047059 were taken twice weekly using calipers. The volume for all measurable tumors wascalculated from the caliper measurements assuming an ovoid shape. The results aresummarized in Figure 3.

[0227] As shown in Figure 3, significant tumor growth inhibition wasobserved with mAb 7.159.2 following twice weekly i.p. administration of l.Omgantibody/mouse. No significant weight loss was observed in any of the groups ofanimals. EXAMPLE 20

INHIBITION OF TUMOR CELL GROWTH IN HUMAN PATIENTS

[0228] A group of human cancer patients diagnosed with pancreatic cancer israndomized into treatment groups. Each patient group is treated with weekly intravenousinjections of mAb 7.159.2, 7.34.1 or 7.251.3 described herein. Each patient is dosed withan effective amount of the antibody ranging from 50 mg/kg to 2,250 mg/kg for 4-8weeks. A control group is given only the standard chemotherapeutic regimen.

[0229] At periodic times during and after the treatment regimen, tumor burdenis assessed by magnetic resonance imaging (MRI). It is found that the patients who havereceived weekly antibody treatment with mAb 7.159.2, 7.34.1 or 7.251.3 show significantreductions in tumor size, compared to patients that do not receive antibody treatment. Insome treated patients, the tumors are no longer detectable. In contrast, tumor sizeincreases or remains substantially the same in the control group. EXAMPLE 21

INHIBITION OF TUMOR CELL GROWTH IN A HUMAN PATIENT

[0230] A human patient is diagnosed with a malignant tumor. The patient istreated with weekly intravenous injections of mAb 7.159.2 for 8 weeks. At periodictimes during and after the treatment regimen, tumor burden is assessed by magneticresonance imaging (MRI). Significant reductions in tumor size are found. EXAMPLE 22

TREATMENT OF ACROMEGALY IN A HUMAN PATIENT

[0231] An adult male is diagnosed with acromegaly. The patient is treatedwith bi-weekly intravenous injections of mAb 7.34.1 over a period of 2( years. As aresult, the patient experiences a significant reduction in the symptoms of acromegaly. 75 WO 2007/070432 PCT/US2006/047059 EXAMPLE 23 TREATMENT OF PSORIASIS IN A HUMAN PATIENT[0232] An adult female is diagnosed with severe psoriasis. The patient is treated with bi-weekly intravenous injections of mAb 7.251.3 over a period of 3 weeks.As a result, the patient experiences a significant reduction in the symptoms of psoriasis. EXAMPLE 24 TREATMENT OF OSTEOPOROSIS IN A HUMAN PATIENT[0233] An adult female is diagnosed with osteoporosis. The patient is treated with bi-weekly intravenous injections of mAb 7.159.2 over a period of a year. As aresult, there is a significant reduction in loss of bone density. EXAMPLE 25 •TREATMENT OF ATHEROSCLEROSIS IN A HUMAN PATIENT[0234] An adult male is diagnosed with atherosclerosis. The patient is treated with bi-weekly intravenous injections of mAb 7.34.1 over a period of a year. As a result,the patient experiences a reduction in the symptoms of atherosclerosis, such as anginapectoris. EXAMPLE 26 TREATMENT OF RESTENOSIS IN A HUMAN PATIENT[0235] An adult male receives angioplasty to relieve a blocked artery.

Following the angioplasty procedure, the patient is treated with bi-weekly intravenousinjections of mAb 7.251.3 over a period of a year. As a result, the patient does notexperience restenosis of the treated artery. EXAMPLE 27 TREATMENT OF DIABETES IN A HUMAN PATIENT[0236] An adult female is diagnosed with diabetes. The patient is treated with bi-weekly intravenous injections of mAb 7.159.2 over a period of a year. As a result, thesymptoms of diabetes are reduced. 76 191718/2

SEQUENCES

[0237] The sub-cloned hybridomas were sequenced to determine their primarystructure at both the nucleotide and amino acid level for both the variable heavy and thevariable light chain genes. The nucleotide and polypeptide sequences of the variableregions of the monoclonal antibodies against IGF-I and IGF-II, as listed in Table 1 , areprovided in the Sequence Listing.

EQUIVALENTS

[0238] The foregoing written specification is considered to be sufficient toenable one skilled in the art to practice the invention. The foregoing description andExamples detail certain preferred embodiments of the invention and describes the bestmode contemplated by the inventors. It will be appreciated, however, that no matter howdetailed the foregoing may appear in text, the invention may be practiced in many waysand the invention should be construed in accordance with the appended claims and anyequivalents thereof. 77 P/15804/125647/596826/l 191718/3

ABXAZ 004VPC (2) .TXTSEQUENCE LISTING <110> Raeber, Olivia

Gazit-Bornstein, GadiYang, XiaodorigCartridge, Susan AnnTonge, David william

<120> BINDING PROTEINS SPECIFIC FOR

INSULIN-LIKE GROWTH FACTORS AND USES THEREOF

<130> ABXAZ.004VPC <160> 102 <170> FastsEQ for windows Version 4.0 <210> 1 <211> 372

<212> DNA <213> Human <400> 1 cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60 acctgcactg tctctggtgg ctccatcagg agtagtagtt actactgggg ctggatccgc 120 cagcccccag ggaaggggct ggagtggatt gggggtatct attatagtgg gagcacctac 180 tacaacccgt ctctcaagag tcgagtcacc atgtccgtag acacgtccaa gaaccagttc 240 tccctgaagc tgagctccgt gaccgccgca gacacggctg tgtattactg tgcgagacaa 300 aggggtcata gcagtggctg gtggtacttc gatctctggg gccgtggcac cctggtcact 360gtctcctcag cc 372 <210> 2<211> 124<212> PRT<213> Human<400> 2

Gin Leu Gin Leu Gin Glu Ser Gly Pro Gly Leu val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser lie Arg Ser Ser 20 25 30 Ser Tyr Tyr Trp Gly Trp lie Arg Gin Pro pro Gly Lys Gly Leu Glu 35 40 45 Trp He Gly Gly lie Tyr Tyr ser Gly Ser Thr Tyr Tyr Asn Pro ser 50 55 60 Leu .Lys Ser Arg val Thr Met ser val Asp Thr Ser Lys Asn Gin Phe 65 70 75 80 ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Al a val Tyr Tyr 85 90 95 cys Al a Arg Gl n Arg Gly Hi s Ser Ser 1 Gly Trp Trp Tyr Phe1 Ί A Asp Leu Trp Gly Arg JL v MGly Thr Leu val Thr Val Ser ser Ala XXV 115 120 <210> 3 <211> 324

<212> DNA <213> Human <400> 3 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagtgtcacc 60atcacttgtc gggcgagtca gggtattagc agctacttag cctggtatca gcagaaacca 120gggaaagccc ctaaactcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcaatggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 78 191718/3

ABXA2 004VPC (2).TXT gaagattttg caacttacta ttgtcaacag gctaacaatt tcccattcac tttcggccct 300gggaccaaag tggatatcaa acga 324 <210> 4 <211> 108

<212> PRT <213> Human <400> 4

Asp lie Gin Met Thr Gin ser Pro Ser Ser Val Ser Ala Ser Val 15 Gly 1 5 10 ASP Ser val Thr He Thr Cys Arg Al a Ser Gin Gl y lie ser Ser Tyr Ala 20 25 30 Leu Trp Tyr Gl n Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu lie Ala 35 40 45 Tyr Ala Ser ser Leu Gin ser Gly val Pro Ser Arg Phe Ser Gly 50 55 60 Asn Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu; Gl n pro 65 Ala 70 75 80 Glu Asp Phe Thr Tyr Tyr Cys Gin Gin Ala Asn Asn Phe Pro phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp lie Lys Arg 100 105 <210> 5 <211> 363

<212> DNA <213> Human <400> 5 caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtc 60 tcctgcaagg cttctggata caccttcacc agttatgata tcaactgggt gcgacaggcc 120 actggacaag ggcttgagtg gatgggatgg atgaacccta acagtggtaa cacaggctat 180 gcacagaagt tccagggcag agtcaccatg accaggaaca cctccataag cacagcctac 240 atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagaccct 300 tactactact actacggtat ggacgtctgg ggccaaggga ccacggtcac cgtctcctca 360 gcc 363 <210> 6 <211> 121

<212> PRT <213> Human <400> 6

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gl y Ala 1 5 10 15 Ser val Lys Val Ser cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Asp lie Asn Trp val Arg Gin Ala Thr Gly Gin Gly Leu Glu Trp Met 35 40 45 Gly Trp Met Asn Pro Asn Ser Gly Asn Thr Gly Tyr Ala Gin Lys Phe 50 55 60 Gin Gly Arg Val Thr Met Thr Arg Asn Thr Ser lie Ser Thr Ala Tyr 65 70 75 80 Met Gl u Leu Ser Ser Leu Arg Ser Glu Asp Thr Al a val Tyr Tyr cys 85 90 95 Ala Arg Asp Pro Tyr Tyr Tyr Tyr Tyr Gly Met ASp val Trp Gly Gl n 100 105 110 Gly Thr Thr Val Thr val ser Ser Ala 115 120 <210> 7<211> 336<212> DNA<213> Human 79 191718/3

ABXAZ 004VPC ¢2).TXT <400> 7 cagtctgtgt tgacgcagcc gccctcagtc tctgcggccc caggacagaa ggtcaccatc 60tcctgctctg gaagcagctc caacattgag aataatcatg tatcctggta ccagcagctc 120ccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 180gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 240actggggacg aggccgatta ttactgcgaa acatgggata ccagcctgag tgctggccgg 500gtattcggcg gagggaccaa gctgaccgtc ctaggt 336 <210> 8 <211> 112

<212> PRT <213> Human <400> 8

Gin 1 ser Val Leu Thr 5 Gin Pro Pro Ser Val10 Ser Ala Ala Pro Gly 15 Gin Lys Val Thr lie Ser Cys Ser Gly Ser Ser Ser Asn lie Glu Asn Asn Hi s val 20 25 30 Ser Trp Tyr Gl n Gin Leu Pro Gly Thr Ala pro Lys Leu Leu Π e 35 40 · 45 Tyr ASP Asn Asn Lys Arg pro ser Gly lie Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly He Thr Gly Leu Gin 65 70 75 80 Thr Gly Asp Glu Al a Asp Tyr Tyr cys Glu Thr Trp Asp Thr Ser Leu 85 90 95 Ser Ala Gly Arg val Phe Gly Gly Gly Thr Lys Leu Thr val Leu Gly 100 105 110 <210> 9 <211> 360

<212> DNA <213> Human <400> 9 caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60 acctgcactg tctctggtgg ctccatcagt agttactact ggagctggat ccggcagccc 120 ccagggaggg gactggagtg gattggctat ttcttctaca gtgggtacac caactacaac 180 ccctccctca agagtcgcgt caccatgtca gttgacacgt ccaagaacca gttctctctg 240 aagctgagct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgtg tataactgga 300 acgacgaagg ggggtatgga cgtctggggc caaggggcca cggtcaccgt ctcctcagcc 360 <210> 10 <211> 120<212> PRT<213> Human 1 1 <400> 10 ) Gin val Gin Leu Gin Glu Ser Gly pro Gly Leu vial Lys Pro Ser Gl u 1 5 10 15 Thr Leu Ser Leu Thr cys Thr val ser Gly Gly Sier lie ser Ser Tyr 20 25 I 30 Tyr Trp ser Trp lie Arg Gin Pro Pro Gly Arg Gly Leu GlU Trp lie 35 40 ί 45 Gly Tyr Phe Phe Tyr Ser Gly Tyr Thr Asn Tyr Asn pro ser Leu Lys 50 55 Thr 60 Asn Gin Phe Ser Arg val Thr Mei: Ser val Asp Ser Lys Ser Leu 65 70 75 , 80 Lys Leu ser Ser val Thr Ala Al a Asp Thr Ala Mal Tyr Tyr Cys Al a 85 90 95 cys lie Thr Gly Thr Thr Lys Gly Gly Met Asp yal Trp Gly Gin Gly 100 105 j 110 Ala Thr val Thr val Ser Ser Ala I 115 120 1 1 80 ,

ABXAZ 004VPC (2) .TXT <210> 11 <211> 336

<212> DNA <213> Human <400> 11 cagtctgtgc tgacgcaggc gccctcagtg tctggggccctcctgcactg ggagaagttc caacatcggg gcaggttaxgtttccaggaa cagcccccaa actcctcatc tatggtaacacctgaccgat tctctggctc caagtctggc acctcagcctcaggctgagg atgaggctga ttattactgc cagtcctatggtattcggcg gagggaccaa gctgaccgtc ctaggt cagggcagag atgtacactg gcaatcggcc ccctggccat acagcagtct ggtcaccatc 60gtaccagcag 120ctcaggggtc 180cactgggctc 240gagtggttcg 300 336 <210> 12<21I> 112<212> PRT<213> Human <400> 12

Gin ser1 val Leu Thr 5 Gin Ala Pro Ser val 10 Ser Gly Ala Pro Gly15 Gl n Arg Val Thr lie ser Cys Thr Gly Arg ser Ser Asn lie Gly Ala Gly 20 25 30 Tyr ASp val His Trp Tyr Gin Gin Phe Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu He Tyr Gly Asn Ser Asn Arg Pro Ser Gly val Pro Asp Arg phe 50 55 60 Ser Gly Ser Lys Ser Gly Thr ser Ala Ser Leu Ala He Thr Gly Leu 65 70 75 80 Gin Ala Glu ASp Glu Ala Asp Tyr Tyr cys Gin ser Tyr Asp Ser Ser 85 90 95 Leu Ser Gly ser val phe Gly Gly Gly Thr Lys Leu Thr val Leu Gly 100 105 110 <210> 13 <211> 360

<212> DNA <213> Human <400> 13 caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60 acctgcactg tctctggtgg ctccatcagt agttactact ggagctggat ccggcagccc 120 ccagggaagg gactggagtg gattgggtat ttcttttaca gtgggtacac caactacaac 180 ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240 aagctgagct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgtg tataacrgga 300 acgacgaagg ggggtatgga cgtctggggc caagggacca cggtcaccgt ctcctcagcc 360 <210> 14<211> 120<212> PRT<213> Human <400> 14

Gin Val Gin Leu Gin Glu Ser Gly Pro Gly Leu Val. Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr val Ser Gly Gly Ser He ser ser Tyr 20 25 30 He Tyr Trp Ser Trp Xie Arg Gin Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Gly Tyr Phe Phe Tyr Ser Gly Tyr Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60 ser Arg val Thr lie ser val Asp Thr Ser Lys Asn Gin Phe Ser Leu 65 70 75 80 Lys Leu Ser Ser val Thr Ala Ala Asp Thr Al a Val Tyr Tyr Cys Ala 85 90 95 Cys He Thr Gly Thr Thr Lys Gly Gly Met Asp Val Trp Gly Gin Gly 81 191718/3

ABXAZ 004VPC (2).TXT 100 105 110

Thr Thr val Thr val Ser Ser Ala 115 120 <210> 15 <211> 336

<212> DNA <213> Human <4Q0> 15 cagtccgtac tgacgcagcc gccctcagtg tctggggccc cagggcagag ggtcaccatc 60tcctgcactg ggagcagctc caacatcggg gcaggttatg atgtacactg gtaccagcag 120cttccaggaa cagcccccaa gctcctcatc tatggtaaca acaatcggcc ctcaggggtc 180cctgaccgat tctctggctc caagtctggc acctcagcct ccctggccat cactgggctc 240caggctgatg atgaggctga ttattactgc cagtcctttg acagcagtct gagtggttcg 300gtattcggcg gagggaccaa gctgaccgtc ctaggt 336 <210> 16 <211> 112

<212> PRT <213> Human <400> 16

Gin Ser Val Leu Thr Gin Pro Pro Ser Val Ser Gly Ala Pro Gly Gl n 1 5 10 15 Arg val Thr lie ser Cys Thr Gly Ser Ser Ser Asn lie Gly Ala Gly 20 25 30 Tyr Asp val His Trp Tyr Gin Gin Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu lie Tyr Gly Asn Asn Asn Arg Pro Ser Gly val Pro Asp Arg Phe 50 55 60 ser Gly Ser Lys ser Gly Thr Ser Ala Ser Leu Ala lie Thr Gly Leu 65 70 75 80 Gin Al a Asp Asp Glu Ala Asp Tyr Tyr Cys Gin Ser Phe Asp Ser Ser 85 90 95 Leu Ser Gly ser Val Phe Gly Gly Gly Thr Lys Leu Thr val Leu Gly 100 105 110 <210> 17 <211> 372

<212> DNA <213> Human <400> 17 cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccatcaac agtagtagta actactgggg ctggatccgc 120cagcccccag ggaagggact ggcgtggatt gggggcatct attatagtgg gagcacctac 180tacaacccgt ccctcaggag tcgagtcacc atgtccgtag acacgtccaa gaaccagttc 240tccctgaagc tgagctctgt gaccgccgca gacacggctg tatattactg tgcgagacaa 300aggggtcata gcagtggctg gtggtacttc gatctctggg gccgtggcac cctggtcact 360gtctcctcag cc 372 <210> 18<211> 124<212> PRT<213> Human<400> 18

Gin Leu Gl n Leu Gin Glu Ser Gly Pro Gly Leu Val Lys pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr val Ser Gly Gly Ser lie Asn Ser Ser 20 25 30 Ser Asn Tyr Trp Gly Trp lie Arg Gin Pro Pro Gly Lys Gly Leu Al a 35 40 45 Trp lie Gly Gly lie Tyr Tyr ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 55 60 82 191718/3

ABXAZ 004VPC C2).TXT

Leu Arg ser Arg Val Thr Met Ser val Asp Thr ser Lys Asn Gin Phe 65 70 75 80

Ser Leu Lys Leu ser ser Val Thr Ala Ala Asp Thr Ala val Tyr Tyr 85 90 95

Cys Ala Arg Gin Arg Gly His ser Ser Gly Trp Trp Tyr Phe Asp Leu 100 105 110

Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Ala 115 120 <210> 19 <211> 324

<212> DNA <213> Human <400> 19 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtcacc 60 atcacttgtc gggcgagtcg gggtattagc agctggttag cctggtatca gcagagacca 120 gggaaagccc ctaagctcct garctatact gcatccagtt tgcaaagtgg ggtcccatca 180 aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240 gaagattttg caacttacta ttgtcaacag gctaacagtt tcccattcac tttcggccct 300 gggaccaaag tggatatcaa acga 324 <210> 20 <211> 108

<212> PRT <213> Human <400> 20 ASP Π e Gin Met Thr Gin Ser Pro Ser Ser val ser Ala ser val Gly 1 5 10 15 Asp Arg Val Thr lie Thr cys Arg Ala Ser Arg Gly lie Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gin Gin Arg Pro Gly Lys Al a Pro Lys Leu Leu lie 35 40 45 Tyr Thr Al a Ser Ser Leu Gin Ser Gly val Pro Ser Arg Phe ser Gly 50 55 60 Ser Gly ser Gly Thr ASp Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Ala Asn Ser phe Pro Phe 85 90 95 Thr Phe Gly pro Gly Thr Lys val ASp He Lys Arg 100 105 <210> 21 <211> 5 <212> PRT <213> Human <400> 21 Ser Tyr Tyr <210> 22 <211> 16

<212> PRT <213> Human <400> 22

Tyr Phe Phe Tyr ser Gly Tyr Thr Asn1 5

Tyr Asn Pro Ser Leu10

Lys Ser15

<210> 23<211> 11<212> PRT 83 191718 <213> Human

ABXAZ 004VPC C2).TXT <400> 23 lie Thr Gly Thr Thr Lys Gly Gly Met Asp val15 10

<210> 24 <211> 14 <212> PRT <213> Human <400> 24 Thr Gly Ser 1 <210> 25 <211> 7 <212> PRT <213> Human <400> 25 Gly Asn Asn 1 <210> 26 <211> 11 <212> PRT <213> Human <400> 26 Gin Ser Phe 1 <210> 27 <211> 5 <212> PRT <213> Human <400> 27 ser Tyr Tyr 1 <210> 28 <211> 16 <212> PRT <213> Human <400> 28 Tyr Phe Phe 1 <210> 29 <211> 11 <212> PRT 10 10 10 15 <213> Human <400> 29 lie Thr Gly Thr Thr Lys Gly Gly Met Asp val15 10 <210> 30 84 191718

„„„ ABXAZ 004VPC C2).TXT <211> 14<212> prt<213> Human<400> 30

Thr Gly Arg Ser Ser Asn lie Gly Ala Gly Tyr Asp Val His 15 10 <210> 31 <211> 7

<212> PRT <213> Human <400> 31

Gly Asn ser Asn Arg Pro ser1 5 <210> 32 <211> 11

<212> PRT <213> Human <400> 32

Gin Ser Tyr Asp Ser ser Leu Ser Gly Ser val15 10 <210> <211> <212> <213> 33 5· PRT Human <400> 33ser Tyr Asp 1 lie Asn5 <210> 34 <211> 17 <212> PRT <213> Human <400> 34

Trp Met Asn1

Gly

Pro Asn5 ser

Gly Asn Thr

Gly Tyr Ala Gin Lys10

Phe Gin15 <210> 35 <211> 11

<212> PRT <213> Human <400> 35

Asp Pro Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val15 10 <210> 36 <211> 13

<212> PRT <213> Human <400> 36

Ser Gly Ser ser Ser Asn lie Glu Asn Asn His val Ser 85 191718 ABXAZ 004VPC C2).TXT15 10 <210> 37 <211> 7

<212> PRT <213>- Human <400> 37

Asp Asn Asn Lys Arg Pro Ser <210> 3δ <211> 12

<212> PRT <213> Human <400> 38

Glu Thr Trp Asp Thr Ser Leu Ser Ala Gly Arg Val1 5 10 <210> 39 <211> 594

<212> DNA <213> Human <400> 39 accatgaaac atctgtggxt cxxccxccxg cxggxggcgg ctcccagatg ggtcctgtcc 60 cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 120 acctgcactg tctctggtgg ctccatcagg agtagtagtr actactgggg ctggatccgc 180 cagcccccag ggaaggggct ggagtggatt gggggtatct attatagtgg gagcacctac 240 tacaacccgt ctctcaagag tcgagtcacc atgtccgtag acacgtccaa gaaccagttc 300 tccctgaagc tgagctccgt gaccgccgca gacacggctg tgtattactg tgcgagacaa 360 aggggtcata gcagtggctg gtggtacttc gatctctggg gccgtggcac cctggtcact 420 gtctcctcag cctccaccaa gggcccaxcg gtcttccccc tggcgccctg ctccaggagc 480 acctccgaga gcacagcggc cctgggctgc ctggtcaagg actacttccc cgaaccggxg 540 acggtgtcgt ggaacxcagg cgctctgacc agcggcgtgc acaccxxccc agcx 594 <210> 40 <211> 198

<212> PRT <213> Human <400> 40

Thr Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Al a Ala Pro Arg 1 5 10 15 Trp val Leu ser Gin Leu Gin Leu Gin Glu Ser Gly Pro Gly Leu val 20 25 30 Lys pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val ser Gly Gly Ser 35 40 45 lie Arg Ser Ser Ser Tyr Tyr Trp Gly Trp lie Arg Gin Pro Pro Gly 50 55 60 Lys Gly Leu Glu Trp lie Gly Gly lie Tyr Tyr Ser Gly Ser Thr Tyr 65 70 75 80 Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Met Ser val Asp Thr Ser 85 90 95 Lys Asn Gin Phe Ser Leu Lys Leu Ser ser Val Thr Ala Ala ASp Thr 100 105 110 Ala val Tyr Tyr cys Ala Arg Gin Arg Gly Hi s Ser ser Gly Trp Trp 115 120 125 Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val ser ser Ala 130 135 140 ser Thr Lys Gly Pro Ser Val Phe Pro Leu a! a Pro Cys Ser Arg Ser 145 150 155 160 Thr Ser Glu ser Thr Ala Al a Leu Gl y cys Leu val Lys Asp Tyr Phe 165 170 175 86 191718

n ABXAZ 004VPC <2).TXT

Pro Glu Pro Val Thr Val ser Trp Asn Ser Gly Ala Leu Thr ser Gly n . 180 185 190

Val His Thr Phe Pro Ala 195 <210> 41 <211> 419

<212> DNA <213> Human <400> 41 atgagggtcc ctgctcagct cctggggctc ctgctgctct ggttcccagg ttccagatgc 60 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagtgtcacc 120 atcacttgtc gggcgagtca gggtattagc agctacttag cctggtatca gcagaaacca 180 gggaaagccc ctaaactcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 240 aggttcagcg gcaatggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 300 gaagattttg caacttacta ttgtcaacag gctaacaatt tcccattcac tttcggccct 360 gggaccaaag tggatatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcc 419 <210> 42 <211> 139

<212> PRT <213> Human <400> 42

Met Arg Val Pro Ala Gl π Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro 1 5 10 15 Gly ser Arg Cys ASp lie Gin Met Thr Gin Ser Pro Ser Ser val Ser 20 25 30 Al a Ser val Gly Asp Ser val Thr lie Thr Cys Arg Ala Ser Gin Gly 35 40 45 lie Ser Ser Tyr Leu Ala Trp Tyr Gin Gl n Lys Pro Gly Lys Ala pro 50 . 55 60 Lys Leu Leu Tie Tyr Ala Al a ser Ser Leu Gl n Ser Gly val Pro Ser 65 70 75 80 Arg Phe Ser Gly Asn Gly ser Gly Thr Asp Phe Thr Leu Thr Tie ser 85 90 95 Ser Leu Gin Pro Glu ASP Phe Ala Thr Tyr Tyr cys Gin Gin Ala Asn 100 105 110 Asn Phe Pro Phe Thr Phe Gly pro Gly Thr Lys val Asp lie Lys Arg 115 120 125 Thr val Al a Ala Pro Ser val Phe He Phe Pro 130 135 <210> 43 <211> 437

<212> DNA <213> Human <400> 43 accatggact ggacctggag gatcctcttc ttggtggcag cagctacaag tgcccactcc 60 caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtc 120 tcctgcaagg cttctggata caccttcacc agttatgata tcaactgggt gcgacaggcc 180 actggacaag ggcttgagtg gatgggatgg atgaacccta acagtggtaa cacaggctat 240 gcacagaagt tccagggcag agtcaccatg accaggaaca cctccataag cacagcctac 300 atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagaccct 360 tactactact actacggtat ggacgtctgg ggccaaggga ccacggtcac cgtctcctca 420 gcctccacca agggccc 437 <210> 44 <211> 145

<212> PRT <213> Human <400> 44

Thr Met: Asp Trp Thr Trp Arg lie Leu Phe Leu Val Ala Ala Ala Thr 87 191718 A BXAZ 004 VPC (2). TXT 1 Ala Hi s 5 10 15 Ser Ser Gl n Val Gin Leu val Gin ser Gly Ala Gl u val Lys Gly 20 25 30 Lys Pro Al a Ser val Lys val Ser cys Lys Ala Ser Gly Tyr Thr Phe Thr 35 40 45 Ser Tyr Asp lie Asn Trp Val Arg Gin Al a Thr Gly Gl n Gly 50 55 60 Leu Glu Trp Met Gly Trp Met Asn Pro Asn Ser Gly Asn Thr Gly Tyr 65 Gin 70 75 80 Ala Lys Phe Gin Gl y Arg val Thr Met Thr Arg Asn Thr Ser lie Thr Ala 85 90 95 Ser Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Al a Val 100 Ala 105 110 Tyr Tyr Cys Arg Asp Pro Tyr Tyr Tyr Tyr Tyr Gly Met ASP 115 120 125 val Trp Gly Gin Gly Thr Thr Val Thr val Ser Ser Ala Ser Thr Lys 130 135 140

Gly 145 <210> 45 <211> 460

<212> DNA <213> Human <400> 45 atggcctggt ctcctctcct cctcaccctt ctcattcact gcacagggtc ctgggcccag 60tctgtgttga cgcagccgcc ctcagtctct gcggccccag gacagaaggt caccatctcc 120tgctctggaa gcagctccaa cattgagaat aatcatgtat cctggtacca gcagctccca 180ggaacagccc ccaaactcct catttatgac aataataagc gaccctcagg gattcctgac 240cgattctctg gctccaagtc tggcacgtca gccaccctgg gcatcaccgg actccagact 300ggggacgagg ccgattatta ctgcgaaaca tgggatacca gcctgagtgc tggccgggta 360rtcggcggag ggaccaagct gaccgtccta ggtcagccca aggctgcccc ctcggtcact 420ctgttcccac cctcctctga ggagctccaa gccaacaagg 460 <210> 46<211> 153<212> PRT<213> Human<400> 46

Met Ala Trp Ser Pro Leu Leu Leu Thr Leu Leu lie His Cys Thr Gly 1 5 10 15 Ala Ser Trp Ala Gin ser val Leu Thr Gin Pro pro Ser val ser Ala 20 25 30 Pro Gly Gin Lys Val Thr He Ser Cys Ser Gly Ser Ser Ser Asn lie 35 40 45 Glu Asn Asn Hi s val Ser Trp Tyr Gin Gin Leu Pro Gly Thr Ala Pro 50 55 60 lie Lys Leu Leu lie Tyr Asp Asn Asn Lys Arg Pro Ser Gly Pro Asp 65 70 75 80 Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly lie Thr 85 90 95 Gly Leu Gin Thr Gly Asp Glu Ala Asp Tyr Tyr cys Glu Thr Trp Asp 100 105 110 Thr Ser Leu ser Al a Gl y Arg val Phe Gly Gly Gly Thr Lys Leu Thr 115 120 125 Val Leu Gly Gin Pro Lys Ala Ala Pro Ser val Thr Leu Phe Pro Pro 130 135 140 Ser Ser Glu Glu Leu Gin Ala Asn Lys 145 150 <210> 47 <211> 613

<212> DNA <213> Human 88 191718

ABXAZ 004VPC (2)-TXT <400> 47 accatgaaac atctgtggtt cttccttctc ctggtggcag ctcccagatg ggtcctgtcc 60 caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 120 acctgcactg tctctggtgg ctccatcagt agttactact ggagctggat ccggcagccc 180 ccagggaggg gactggagtg gattggctat ttcttttaca gtgggtacac caactacaac 240 ccctccctca agagtcgcgt caccatgtca gttgacacgt ccaagaacca gttctctctg 300 aagctgagct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgtg tataactgga 360 acgacgaagg ggggtatgga cgtctggggc caaggggcca cggtcaccgt ctcctcagcc 420 tccaccaagg gcccatcggt cttccccctg gcgccctgct ccaggagcac ctccgagagc 480 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 540 aactcaggcg ctctgaccag cggcgtgcac accttcccag ctgtcctaca gtcctcagga 600 ctctactccc tea 613 <210> 48<211> 204<212> PRT<213> Human<400> 48

Thr Met Lys His Leu Trp Phe Phe Leu Leu Leu val Ala Ala Pro Arg 1 5 10 15 Trp Val Leu Ser Gin val Gin Leu Gin Glu ser Gly Pro Gly Leu val 20 25 30 Lys Pro ser Glu Thr Leu Ser Leu Thr cys Thr Val Ser Gly Gly ser 35 40 45 lie Ser Ser Tyr Tyr Trp Ser Trp lie Arg Gin pro Pro Gly Arg Gly 50 55 60 Leu Gl u Trp lie Gly Tyr Phe Phe Tyr Ser Gly Tyr Thr Asn Tyr Asn 65 70 75 80 Pro Ser Leu Lys Ser Arg val Thr Met Ser Val Asp Thr ser Lys Asn 85 90 95 Gin Phe Ser Leu Lys Leu Ser ser Val Thr Ala Ala ASp Thr Ala val 100 105 110 Tyr Tyr cys Ala Cys lie Thr Gly Thr Thr Lys Gly Gly Met Asp Val 115 120 125 Trp Gly Gin Gly Ala Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 130 135 140 Gl u Pro ser val Phe Pro Leu Al a Pro cys Ser Arg ser Thr ser Ser 145 150 155 160 Thr Ala Ala Leu Gly cys Leu Val Lys Asp Tyr Phe Pro Glu Pro val 165 170 175 Thr val ser Trp Asn ser Gly Ala Leu Thr Ser Gly val Hi s Thr Phe 180 185 190 pro Ala Val Leu Gin ser Ser Gly Leu Tyr ser Leu 195 200 <210> 49 <211> 432

<212> DNA <213> Human <400> 49 cctctgctcc tcactctcct cgctcactgc acagggtcct gggcccagtc tgtgctgacg 60 caggcgccct cagtgtctgg ggccccaggg cagagggtca ccatctcctg cactgggaga 120 agttccaaca tcggggcagg ttatgatgta cactggtacc agcagtttcc aggaacagcc 180 cccaaactcc teatetatgg taacagcaat cggccctcag gggtccctga ccgattctct 240 ggctccaagt ctggcacctc agcctccctg gccatcactg ggctccaggc tgaggatgag 300 getgattatt actgccagtc ctatgacagc agtctgagtg gttcggtatt eggeggaggg 360 accaagctga ccgtcctagg tcagcccaag gctgccccct cggtcactct gttcccgccc 420 tcctctgagg ag 432 <210> 50 <211> 144

<212> PRT <213> Human 89 191718

ABXAZ 004VPC (2).TXT

Pro Leu Leu Leu Thr Leu Leu Ala Hi s Cys Thr Gly Ser Trp Ala .Gin 1 val Thr 5 10 15 Ser Leu Gin Al a Pro ser Val Ser Gly Ala Pro Gly Gin Arg val Thr 20 25 30 lie Ser Cys Thr Gly Arg Ser Ser Asn He Gly Ala Gl y Tyr val 3 5 40 45 Asp Hi s Trp Tyr Gin Gin Phe Pro Gly Thr Ala Pro Lys Leu Leu lie 50 Gly 55 60 Tyr Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 65 70 75 80 Gly ser Lys Ser Gly Thr Ser Ala Ser Leu Ala lie Thr Gly Leu Gin Ala Glu Glu 85 90 95 Asp Ala Asp Tyr Tyr Cys Gin ser Tyr ASp ser Ser Leu 100 105 110 Ser Gly Ser Val Phe Gly Gly Gly Thr Lys Leu Thr val Leu Gly Gin 115 Ala 120 125 Pro Lys Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 130 135 140 <210> 51 <211> 543

<212> DNA <213> Human <400> 51 atgaagcatc tgtggttctt ccttctcctg gtggcagctc ccagatgggt cctgtcccag 60gtgcagctgc aggagtcggg cccaggactg gtgaagcctt cggagaccct gtccctcacc 120tgcactgtct ctggtggctc catcagtagt tactactgga gctggatccg gcagccccca 180gggaagggac tggagtggat tgggtatttc ttttacagtg ggtacaccaa ctacaacccc 240tccctcaaga gtcgagtcac catatcagta gacacgtcca agaaccagtt ctccctgaag 300ctgagctctg tgaccgctgc ggacacggcc gtgtattact gtgcgtgtat aactggaacg 360acgaaggggg gtatggacgt ctggggccaa gggaccacgg tcaccgtctc ctcagcctcc 420accaagggcc catcggtctt ccccctggcg ccctgctcca ggagcacctc cgagagcaca 480gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 540tea 543 <210> 52 <211> 181

<212> PRT <213> Human <400> 52

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp 1 5 10 15 val Leu Ser Gin Val Gin Leu Gin Glu ser Gly Pro Gly Leu Val Lys 20 25 30 Pro ser Glu Thr Leu Ser Leu Thr cys Thr val Ser Gly Gly Ser lie 35 40 45 ser Ser Tyr Tyr Trp Ser Trp lie Arg Gin Pro Pro Gly Lys Gly Leu 50 55 60 Glu Trp lie Gly Tyr Phe Phe Tyr ser Gly Tyr Thr Asn Tyr Asn Pro 65 70 75 80 ser Leu Lys Ser Arg val Thr lie ser val ASp Thr ser Lys Asn Gin 85 90 95 Phe Ser Leu Lys Leu ser ser val Thr Ala Ala Asp Thr Ala Val Tyr 100 105 110 Tyr Cys Ala Cys He Thr Gly Thr Thr Lys Gly Gly Met Asp val Trp 115 120 125 Gly Gin Gly Thr Thr val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 130 135 140 Ser Val Phe Pro Leu Ala Pro cys ser Arg Ser Thr Ser Glu Ser Thr 145 150 155 160 Ala Al a Leu Gly cys Leu val Lys Asp Tyr Phe Pro Glu Pro Val Thr 165 170 175 val Ser Trp Asn Ser 180 90 191718 ABXAZ 004VPC ζ2).ΤΧΤ <210> 53 <211> 451

<212> DNA <213> Human <400> 53 tcctctgctc ctcactctcc tcgctcactg cacagggtcc tgggcccagt ctgtactgac 60 gcagccgccc tcagtgtctg gggccccagg gcagagggtc accatctcct gcactgggag 120 cagctccaac atcggggcag gttatgatgt acactggtac cagcagcttc caggaacagc 180 ccccaagctc ctcatctatg gtaacaacaa tcggccctca ggggtccctg accgatrctc 240 tggctccaag tctggcacct cagcctccct ggccatcact gggctccagg ctgatgatga 300 ggctgattat tactgccagt cctttgacag cagtctgagt ggttcggtat tcggcggagg 360 gaccaagctg accgtcctag gtcagcccaa ggctgccccc tcggtcactc tgttcccgcc 420 ctcctctgag gagcrccaag ccaacaagga a 451 <210> 54 <211> 145

<212> PRT <213> Human <400> 54

Pro Leu Leu Leu Thr Leu Leu Ala His Cys Thr Gly Ser Trp Al a Gin 1 5 10 15 Ser val Leu Thr Gin Pro Pro Ser val Ser Gly Al a Pro Gly Gin Arg 20 25 30 val Thr lie Ser cys Thr Gly Ser Ser Ser Asn lie Gly Ala Gly Tyr 35 40 45 Asp Val Hi s Trp Tyr Gin Gl n Leu Pro Gly Thr Ala Pro Lys Leu Leu 50 55 60 lie Tyr Gly Asn Asn Asn Arg Pro Ser Gly val Pro Asp Arg phe Ser 65 70 75 80 Gly Ser Lys Ser Gly Thr Ser Al a ser Leu Ala lie Thr Gly Leu Gl n 85 90 95 Ala Asp ASp Glu Ala Asp Tyr Tyr Cys Gin Ser Phe ASp Ser Ser Leu 100 105 110 ser Gly Ser val Phe Gly Gly Gly Thr Lys Leu Thr val Leu Gly Gin 115 120 125 Glu Glu Pro Lys Ala Ala Pro ser Val Thr Leu Phe Pro Pro Ser Ser 130 135 140 Leu 145 <210> 55 <211> 559

<212> DNA <213> Human <400> 55 accatgaaac atctgtggtt cttcctcctg ctggtggcgg ctcccagatg ggtcctgtcc 60 cagctgcagc tgcaggagrc gggcccagga ctggtgaagc cttcggagac cctgtccctc 120 acctgcactg tctctggtgg ctccatcaac agtagtagta actactgggg ctggatccgc 180 cagcccccag ggaagggact ggcgtggatt gggggcatct attatagtgg gagcacctac 240 tacaacccgt ccctcaggag tcgagtcacc atgtccgtag acacgtccaa gaaccagttc 300 tccctgaagc tgagctctgt gaccgccgca gacacggctg tatattactg tgcgagacaa 360 aggggtcata gcagtggctg gtggtacttc gatctctggg gccgtggcac cctggtcact 420 grctcctcag cctccaccaa gggcccatcg gtcttccccc tggcgccctg ctccaggagc 480 acctccgaga gcacagcggc cctgggctgc ctggtcaagg actacttccc cgaaccggtg 540 a-cggtgtcgt ggaactcag 559 <210> 56 <211> 186

<212> PRT <213> Human <400> 56 91 191718

, ABXAZ 004VPC (2).TXT

Thr Met Lys ms Leu Trp phe Phe Leu Leu Leu val Ala Ala Pro Arq 1 n 5 10 15

Trp Val Leu Ser Gin Leu Gin Leu Gin Glu ser Gly Pro Gly Leu val 20 25 30

Lys Pro Ser Glu Thr Leu Ser Leu Thr cys Thr Val Ser Gly Gly ser35 40 45

He Asn Ser Ser Ser Asn Tyr Trp Gly Trp lie Arg Gin Pro Pro Gly50 55 60

Lys Gly Leu Ala Trp lie Gly Gly lie Tyr Tyr Ser Gly Ser Thr Tyr65 70 75 80

Tyr Asn Pro Ser Leu 85 Arg Ser Arg Val Thr 90 Met Ser Val Asp Thr 95 Ser Lys Asn Gin Phe Ser Leu Lys Leu Ser Ser val Thr Al a Al a ASp Thr 100 105 110 Al a val Tyr Tyr Cys Ala Arg Gin Arg Gly His Ser Ser Gly Trp T rp 115 120 125 Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser ser Ala 130 135 140 Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser 145 150 155 160 Thr Ser Gl u Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 165 170 175 Pro Glu Pro Val Thr Val Ser Trp Asn Ser 180 185 <210> 57 <211> 532

<212> DNA <213> Human <400> 57 atgagggtcc ccgctcagct cctggggctc ctgctgctct ggttcccagg ttccagatgc 60 gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtcacc 120 atcacttgtc gggcgagtcg gggtattagc agctggttag cctggtatca gcagagacca 180 gggaaagccc ctaagctcct gatctatact gcatccagtt tgcaaagtgg ggtcccatca 240 aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 300 gaagattttg caacttacta ttgtcaacag gctaacagtt tcccattcac tttcggccct 360 gggaccaaag tggatatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 420 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 480 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg ta 532 <210> 58 <211> 177

<212> PRT <213> Human <400> 58

Met Arg val Pro Ala Gin Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro 1 5 10 15 Gly Ser Arg cys ASp lie Gin Met Thr Gin ser Pro Ser Ser val Ser 20 25 30 Ala Ser Val Gly ASp Arg val Thr lie Thr cys Arg Al a Ser Arg Gly 35 40 45 lie Ser Ser Trp Leu Ala Trp Tyr Gl n Gin Arg Pro Gly Lys Al a Pro 50 55 60 Lys Leu Leu lie Tyr Thr Ala Ser Ser Leu Gin Ser Gly val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Xie ser 85 90 95 Ser Leu Gin Pro Glu ASP Phe Ala Thr Tyr Tyr cys Gin Gl n Al a Asn 100 105 110 ser Phe Pro Phe Thr Phe Gly pro Gly Thr Lys Val Asp lie Lys Arg 115 120 125 Thr Val Ala Ala Pro Ser val phe lie Phe Pro pro Ser ASp Glu Gin 130 135 140 Leu Lys ser Gly Thr Ala Ser val val cys Leu Leu Asn Asn Phe Tyr 145 150 155 160 92 191718

ABXAZ 004VPC (2).TXT

Pro Arg Glu Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser165 170 175

Gly <210> 59 <211> 594

<212> DNA <213> Human <400> 59 accatgaaac atctgtggtt cttcctcctg ctggtggcgg ctcccagatg ggtcctgtcc 60 cagctgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 120 acctgcactg tctctggtgg ctccatcagc agtagtagtt actactgggg ctggatccgc 180 cagcccccag ggaaggggct ggagtggatt gggggtatct attatagtgg gagcacctac 240 tacaacccgt ctctcaagag tcgagtcatc atgtccgtag acacgtccaa gaaccagttc 300 tccctgaagc tgagctccgt gaccgccgca gacacggctg tgtattactg tgcgagacaa 360 a-SggQtcata gcagtggctg gtggtacttc gatctctggg gccgtggcac cctggtcact 420 gtctcctcag cctccaccaa gggcccatcg gtcttccccc tggcgccctg ctccaggagc 480 acctccgaga gcacagcggc cctgggctgc ctggtcaagg actacttccc cgaaccggtg 540 acggtgtcgt ggaactcagg cgctctgacc agcggcgtgc acaccttccc agct 594 <210> 60 <211> 198

<212> PRT <213> Human <400> 60

Thr Met lys His Leu Trp Phe Phe Leu Leu Leu val Ala Ala pro Arg 1 5 10 15 Trp val Leu Ser Gin Leu Gin Leu Gin Glu ser Gly Pro Gly Leu val 20 25 30 Lys Pro ser Glu Thr Leu ser Leu Thr Cys Thr val Ser Gly Gly Ser 35 40 45 lie ser Ser Ser Ser Tyr Tyr Trp Gly Trp lie Arg Gin Pro pro Gly 50 55 60 Lys Gly Leu Glu Trp lie Gly Gly lie Tyr Tyr Ser Gly Ser Thr Tyr 65 70 75 80 Tyr Asn Pro ser Leu Lys Ser Arg Val Thr lie Ser val Asp Thr Ser 85 90 95 Lys Asn Gin Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Al a ASp Thr 100 105 110 Ala Val Tyr Tyr cys Ala Arg Gin Arg Gly His Ser Ser Gly Trp Trp 115 120 125 Tyr Phe ASP Leu Trp Gly Arg Gly Thr Leu val Thr val Ser Ser Ala 130 135 ‘140 Ser Thr Lys Gly Pro Ser Val Phe pro Leu Ala pro cys ser Arg ser 145 150 155 160 Thr Ser Glu Ser Thr Ala Ala Leu Gly cys Leu val Lys Asp Tyr Phe 165 170 175 Pro Glu pro Val Thr val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 180 185 190 val His Thr Phe Pro Ala 195 <210> 61 <211> 419

<212> DNA <213> Human <400> 61 atgagggtcc ctgctcagct cctggggctc ctgctgctct ggttcccagg ttccagatgc 60gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtcacc 120atcacttgtc gggcgagtca gggtattagc agctacttag cctggtatca gcagaaacca 180gggaaagccc ctaaactcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 240aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 300 93 191718

ABXAZ 004VPC (2).TXT gaagattttg caacttacta ttgtcaacag gctaacaatt tcccattcac tttcggccct 360gggaccaaag tggatatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcc 419 <210> 62 <211> 139

<212> PRT <213> Human <400> 62

Met Arg Val Pro Ala Gin Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro 1 5 10 15 Gly Ser Arg cys ASp lie Gin Met Thr Gin Ser Pro ser Ser Val Ser Ala 20 25 30 Ser Val Gly ASp Arg Val Thr He Thr Cys Arg Ala Ser Gin Gl y lie 35 40 45 Ser Ser Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ala pro 50 55 60 Lys Leu Leu lie Tyr Ala Ala ser Ser Leu Gin ser Gly val Pro Ser 65 Phe 70 75 80 Arg Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser 85 90 95 ser Leu Gin Pro Gl u Asp Phe Ala Thr Tyr Tyr cys Gin Gin Ala Asn 100 105 110 Asn Phe Pro Phe Thr Phe Gly Pro Gly Thr Lys val Asp lie Lys Arg 115 120 125 Thr val Ala Ala pro Ser Val Phe lie Phe pro 130 135 <210> 63 <211> 437

<212> DNA <213> Human <400> 63 accatggact ggacctggag gatcctcttc ttggtggcag cagctacaag tgcccactcc 60 caggtgcagc tggtgcagtc tggggctgag gtgaagaagc ctggggcctc agtgaaggtc 120 tcctgcaagg cttctggata caccttcacc agttatgata tcaactgggt gcgacaggcc 180 actggacaag ggcttgagtg gatgggatgg atgaacccta acagtggtaa cacaggctat 240 gcacagaagt tccagggcag agrcaccatg accagggaca cctccataag cacagcctac 300 atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc gagagaccct 360 tactactact actacggtat ggacgtctgg ggccaaggga ccacggtcac cgtctcctca 420 gcctccacca agggccc 437 <210> 64 <211> 145

<212> PRT <213> Human <400> 64

Thr Met Asp Trp Thr Trp Arg lie Leu Phe Leu Val Ala Ala Ala Thr 1 5 10 15 Ser Ala Hi s Ser Gin val Gin Leu Val Gin ser Gly Ala Glu Val Lys 20 25 30 Lys Pro Gly Al a ser val Lys val Ser cys Lys Ala Ser Gly Tyr Thr 35 40 45 Phe Thr Ser Tyr Asp lie Asn Trp val Arg Gin Ala Thr Gly Gin Gly 50 55 60 Leu Glu Trp Met Gly Trp Met Asn Pro Asn ser Gly Asn Thr Gly Tyr 65 70 75 80 Ala Gin Lys Phe Gin Gly Arg val Thr Met Thr Arg Asp Thr Ser lie 85 90 95 Ser Thr Ala Tyr Met Gl u Leu ser Ser Leu Arg Ser Glu Asp Thr Ala 100 105 110 val Tyr Tyr Cys Ala Arg Asp pro Tyr Tyr Tyr Tyr Tyr Gly Met Asp 115 120 125 val Trp Gly Gin Gly Thr Thr val Thr val Ser Ser Ala ser Thr Lys 130 135 140 94 191718

, ABXAZ 004VPC (2).TXT

Gly145 <210> 65 <211> 460

<212> DNA <213> Human <400> 65 atggcctggt ctcctctcct cctcaccctt ctcattcact gcacagggtc ctgggcccag 60tctgtgttga cgcagccgcc ctcagtctct gcggccccag gacagaaggt caccatctcc 120tgctctggaa gcagctccaa cattgagaat aatcatgtat cctggtacca gcagctccca 180ggaacagccc ccaaactcct catttatgac aataataagc gaccctcagg gattcctgac 240cgattctctg gctccaagtc tggcacgtca gccaccctgg gcatcaccgg actccagact 300ggggacgagg ccgattatta ctgcgaaaca tgggatacca gcctgagtgc tggccgggta 360ttcggcggag ggaccaagct gaccgtccta ggtcagccca aggctgcccc ctcggtcact 420ctgttcccac cctcctctga ggagctccaa gccaacaagg 460 <210> 66 <211> 153

<212> PRT <213> Human <400> 66

Met Ala Trp Ser Pro Leu Leu Leu Thr Leu Leu lie His Cys Thr Gly 1 5 10 15 Ser Trp Al a Gin Ser val Leu Thr Gin Pro Pro Ser val Ser Ala Al a 20 25 30 Pro Gly Gin Lys Val Thr Xie Ser cys Ser Gly ser Ser Ser Asn lie 35 40 45 Glu Asn Asn His val Ser Trp Tyr Gin Gin Leu Pro Gly Thr Ala Pro 50 55 60 Lys Leu Leu lie Tyr Asp Asn Asn Lys Arg Pro Ser Gly lie pro Asp 65 70 75 80 Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly lie Thr 85 90 95 Gly Leu Gin Thr Gly ASp Gl u Ala ASp Tyr Tyr Cys Glu Thr Trp Asp 100 105 110 Thr Ser Leu Ser Al a Gly Arg Val Phe Gly Gly Gly Thr Lys Leu Thr 115 120 125 val Leu Gly Gin Pro Lys Al a Ala Pro ser val Thr Leu Phe Pro Pro 130 135 140 Ser Ser Glu Glu Leu Gl n Ala Asn Lys 145 150 <210> 67 <211> 613

<212> DNA <213> Human <400> 67 accatgaaac atctgtggtt cttccttctc ctggtggcag ctcccagatg ggtcctgtcc 60 caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 120 acctgcactg tctctggtgg ctccatcagt agttactact ggagctggat ccggcagccc 180 ccagggaagg gactggagtg gattggctat ttcttttaca gtgggtacac caactacaac 240 ccctccctca agagtcgcgt caccatctca gttgacacgt ccaagaacca gttctctctg 300 aagctgagct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgcg tataactgga 360 acgacgaagg ggggtatgga cgtctggggc caagggacca cggtcaccgt ctcctcagcc 420 tccaccaagg gcccatcggt cttccccctg gcgccctgct ccaggagcac ctccgagagc 480 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 540 aactcaggcg ctctgaccag cggcgtgcac accttcccag ctgtcctaca gtcctcagga 600ctctactccc tea 613 <210> 68 <211> 204

<212> PRT 95 191718 <213> Human ABXAZ 004VPC C2). TXT <400> 68 Thr Met Lys His Leu Trp Phe Phe Leu Leu Leu val Ala Ala Pro Arg 1 5 10 15 Trp val Leu Ser Gin val Gin Leu Gin Glu Ser Gly Pro Gly Leu val 20 25 30 Lys Pro Ser Glu Thr Leu ser Leu Thr cys Thr val Ser Gly Gly Ser 35 40 45 lie Ser Ser Tyr Tyr Trp Ser Trp lie Arg Gin Pro Pro Gly Lys Gly 50 55 60 Leu Glu Trp lie Gly Tyr Phe Phe Tyr Ser Gly Tyr Thr Asn Tyr Asn 65 70 75 80 Pro Ser Leu Lys Ser Arg val Thr lie Ser val ASp Thr Ser Lys Asn 85 90 95 Gin Phe ser Leu Lys Leu Ser Ser val Thr Ala Ala Asp Thr Ala val 100 105 110 Tyr Tyr Cys Ala Arg lie Thr Gly Thr Thr Lys Gly Gly Met Asp val 115 120 125 Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala ser Thr Lys Gly 130 135 140 Pro Ser val Phe Pro Leu Ala Pro Cys Ser Arg ser Thr Ser Glu ser 145 150 155 160 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro val 165 170 175 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly val His Thr Phe 180 185 190 Pro Ala val Leu Gin Ser Ser Gly Leu Tyr Ser Leu 195 200 <210> 69 <211> 432

<212> DNA <213> Human <400> 69 cctctgctcc tcactctcct cgctcactgc acagggtcct gggcccagtc tgtgctgacg 60 cagccgccct cagtgtctgg ggccccaggg cagagggtca ccatctcctg cactgggaga 120 agttccaaca tcggggcagg ttatgatgta cactggtacc agcagttgcc aggaacagcc 180 cccaaactcc tcatctatgg taacagcaat cggccctcag gggtccctga ccgattctct 240 ggctccaagt ctggcacctc agcctccctg gccatcactg ggctccaggc tgaggatgag 300 gctgattatt actgccagtc ctatgacagc agtctgagtg gttcggtatt cggcggaggg 360 accaagctga ccgtcctagg tcagcccaag gctgccccct cggtcactct gttcccgccc 420 tcctctgagg ag 432 <210> 70 <211> 144

<212> PRT <213> Human <4Q0> 70 pro Leu Leu Leu Thr Leu Leu Ala Hi S cys Thr Gly Ser Trp Ala Gin 1 5 10 15 Ser Val Leu Thr Gin pro pro Ser Val Ser Gly Ala Pro Gly Gin Arg 20 25 30 Val Thr lie Ser Cys Thr Gly Arg Ser Ser Asn lie Gly Ala Gly Tyr 35 40 Gly. 45 ASp Val Hi s Trp Tyr Gin Gin Leu Pro Thr Ala Pro Lys Leu Leu 50 55 60 lie Tyr Gly Asn Ser Asn Arg Pro Ser Gly val Pro Asp Arg Phe Ser 65 70 75 80 Gly Ser Lys Ser Gly Thr ser Ala Ser Leu Ala lie Thr Gly Leu Gin 85 90 95 Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gin Ser Tyr Asp Ser Ser Leu 100 105 110 ser Gly Ser val Phe Gly Gly Gly Thr Lys Leu Thr val Leu Gly Gin 115 120 125 96 191718

_ ABXAZ 004VPC C2).TXT

Pro Lys Ala Ala Pro ser Val Thr Leu Phe Pro pro ser ser Glu Glu 130 135 140 <210> 71 <211> 543

<212> DNA <213> Human <400> 71 atgaagcatc tgtggttctt ccttctcctg gtggcagctc ccagatgggt cctgtcccag 60 gtgcagctgc aggagtcggg cccaggactg gtgaagcctt cggagaccct gtccctcacc 120 tgcactgtct ctggtggctc catcagtagt tactactgga gctggatccg gcagccccca 180 gggaagggac tggagtggat tgggtatttc ttttacagtg ggtacaccaa ctacaacccc 240 tccctcaaga gtcgagtcac catatcagta gacacgtcca agaaccagtt ctccctgaag 300 ctgagctctg tgaccgctgc ggacacggcc gtgtattact gtgcgcgtat aactggaacg 360 acgaaggggg gtatggacgt ctggggccaa gggaccacgg tcaccgtctc ctcagcctcc 420 accaagggcc catcggtctt ccccctggcg ccctgctcca ggagcacctc cgagagcaca 480 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 540 tea 543 <210> 72 <211> 181

<212> PRT <213> Human <400> 72

Met Lys Hi S Leu Trp Phe Phe Leu Leu Leu Val Ala Ala pro Arg Trp 1 5 10 15 val Leu Ser Gin Val Gin Leu Gin Glu ser Gly pro Gly Leu val Lys 20 25 30 Pro ser Gl u Thr Leu Ser Leu Thr Cys Thr val Ser Gly Gly Ser He 35 40 45 ser ser Tyr Tyr Trp ser Trp lie Arg Gin Pro Pro Gly Lys Gly Leu 50 55 60 Glu Trp lie Gly Tyr Phe Phe Tyr Ser Gly Tyr Thr Asn Tyr Asn Pro 65 70 75 80 ser Leu Lys ser Arg Val Thr He Ser Val ASp Thr Ser Lys Asn Gin 85 90 95 Phe Ser Leu Lys Leu Ser ser Val Thr Ala Ala ASp Thr Ala val Tyr 100 105 110 Tyr cys Ala Arg He Thr Gly Thr Thr Lys Gly Gly Met Asp val Trp 115 120 125 Gly Gin Gly Thr Thr val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 130 135 140 Ser Val Phe Pro Leu Ala Pro Cys Ser Arg ser Thr Ser GlU Ser Thr 145 150 155 160 Ala Ala Leu Gly cys Leu Val Lys Asp Tyr phe Pro Glu Pro val Thr 165 170 175 val Ser Trp Asn Ser 180 <210> 73 <211> 451

<212> DNA <213> Human <400> 73 tcctctgctc ctcactctcc tcgctcactg cacagggtcc tgggcccagt ctgtactgac 60 gcagccgccc tcagtgtctg gggccccagg gcagagggtc accatctcct gcactgggag 120 cagctccaac atcggggcag gttatgatgt acactggtac cagcagcttc caggaacagc 180 ccccaagctc ctcatctatg gtaacaacaa tcggccctca ggggtccctg accgattctc 240 tggctccaag tctggcacct cagcctccct ggccatcact gggctccagg ctgaagatga 300 ggctgattat tactgccagt cctttgacag cagtctgagt ggttcggtat teggeggagg 360 gaccaagctg accgtcctag gtcagcccaa ggctgccccc tcggtcactc tgttcccgcc 420 ctcctctgag gagctccaag ccaacaagga a 451 97 191718 <210> 74<211> 145<212> PRT<213> Human<400> 74

Pro Leu Leu 1 Ser val Leu val Thr He Asp val 35Hi s He 65 Gly 50 Tyr Gly Ser Lys Al a Glu Asp Ser Gly Ser Pro Lys 115 Ala Leu 145 130 A BXAZ 004 VPC (2). TXT Leu Thr Leu Leu Ala Hi s cys Thr Gly Ser Trp Ala Gin Thr 5 10 15 Gin pro pro Ser val Ser Gly Ala Pro Gly Gin Arg 20 Thr 25 30 Ser Cys Gly Ser 40 Ser ser Asn lie Gly Ala Gly Tyr Trp Tyr Gin Gin Leu Pro Gly Thr Ala A3 Pro Lys Leu Leu 55 60 Asn Asn Asn Arg pro ser Gly val Pro Asp Arg Phe ser Gly 70 75 80 Ser Thr ser Ala Ser Leu Ala lie Thr Gly Leu Gin Glu 85 90 95 Al a Asp Tyr Tyr Cys Gin Ser Phe Asp ser Ser Leu 100 105 110 Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gl n Ala 120 125 pro Ser val Thr Leu Phe Pro Pro Ser ser Gl u Glu 135 140 <210> 75<211> 119<212> PRT<213> Human <400> 75

Gin val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 ser Val Lys val Ser Cys Lys Al a Ser Gly Tyr Thr Phe Thr ser Tyr 20 25 30 Asp lie Asn Trp Val Arg Gin Al a Thr Gly Gin Gly Leu Glu Trp Met 35 40 45 Gly Trp Met Asn Pro Asn Ser Gly Asn Thr Gly Tyr Ala Gl n Lys Phe 50 55 60 Gl n Gly Arg val Thr Met Thr Arg Asn Thr ser lie Ser Thr Ala Tyr 65 70 75 Val 80 Met Gl U Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Tyr Tyr Cys 85 90 95 Al a Arg Tyr Tyr Tyr Tyr Tyr Gly Met ASp val Trp Gly Gin Gly Thr 100 105 110 Thr val Thr Val Ser Ser Ala 115 <210> 76<211> 110<212> PRT<213> Human <400> 76

Gl n Ser val Leu Thr Gin pro Pro Ser val ser Ala Ala Pro Gly Gin 1 5 10 15 Lys val Thr lie Ser Cys Ser Gly Ser ser Ser Asn He Gly Asn Asn 20 25 30 Tyr Val Ser Trp Tyr Gin Gl n Leu Pro Gly Thr Ala pro Lys Leu Leu 35 40 45 lie Tyr ASp Asn Asn Lys Arg Pro ser Gly lie Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys ser Gly Thr Ser Al a Thr Leu Gly He Thr Gly Leu Gin 65 70 75 80 98 191718

ABXAZ 004VPC (2).TXT

Thr Gly Asp Gl u Phe 100 Ala Asp85 Tyr Tyr Cys Gly Thr Trp Asp ser90 Ser 95 Leu Ser Ala val Gly Gly Gly Thr Lys 105 Leu Thr val Leu Gly 110 <210> 77<211> 118<212> PRT<213> Human <400> 77 Gin Leu Gin Leu Gin Glu Ser Gly Pro Gly Leu val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr val Ser Gly Gly Ser lie Ser Ser Ser 20 Gly 25 30 Ser Tyr Tyr T rp Trp lie Arg Gl n Pro Pro Gly Lys Gly Leu Gl u Trp He 35 40 45 Gly Ser He Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg val Thr He ser val Asp Thr Ser Lys Α5Π Gin Phe 65 70 75 80 Ser Leu Lys Leu Ser Ser Val Thr Al a Ala ASp Thr Ala val Tyr Tyr Cys Ala 85 90 • 95 Arg Ser Ser Trp Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala 115 <210> 78<211> 108<212> PRT<213> Human <400> 78 Asp lie Gin Met Thr Gl n Ser pro Ser Ser val Ser Ala Ser val Gly 1 5 10 15 Asp Arg val Thr lie Thr cys Arg a! a Ser Gin Gly lie Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu lie 35 40 45 Tyr Ala Ala Ser Ser Leu Gl n Ser Gly val Pro Ser Arg Phe Ser Gl y 50 55 60 Ser Gly Ser Gly Thr ASp Phe Thr Leu Thr lie Ser Ser Leu Gl n Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr cys Gin Gin Ala Asn Ser Phe pro phe 85 90 95 Thr Phe Gly pro Gly Thr Lys val Asp He Lys Arg 100 105 <210> 79<211> 117<212> PRT<213> Human <400> 79

Gin Val Gin Leu Gin Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr val ser Gly Gly Ser lie Ser Ser Tyr 20 25 30 Tyr Trp Ser Trp lie Arg Gin pro Pro Gly Lys Gly Leu Gl u Trp He 35 40 45 Gly Tyr lie Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn pro Ser Leu Lys 50 55 60 Ser Arg val Thr lie ser val Asp Thr Ser Lys Asn Gin Phe Ser Leu 65 70 75 80 99 191718

ABXAZ 004VPC C2).TXT

Lys Leu Ser Ser val 85 Thr Ala Ala Asp Thr Ala val90 Tyr Tyr Cys 95 Al a Arg lie Thr Gly Thr Gly Met Asp val Trp Gly Gin Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala 115 <210> 80<211> 112<212> PRT<213> Human <400> 80

Gin Ser val Leu Thr Gin Pro Pro Ser val ser Gly Ala Pro Gly Gl π 1 5 10 15 Arg val Thr lie Ser Cys Thr Gly Ser Ser Ser Asn lie Gly Ala Gly 20 25 30 Tyr Asp val His Trp Tyr Gin Gin Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu lie Tyr Gly Asn Ser Asn Arg Pro Ser Gly val pro Asp Arg Phe 50 55 60 ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Al a lie Thr Gly Leu 65 70 75 80 Gin Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gin Ser Tyr Asp ser Ser 85 90 95 Leu Ser Gl y Ser Val Phe Gly Gly Gly Thr Lys Leu Thr val Leu Gly 100 105 110 <210> 81 <211> 7

<212> PRT <213> Human <400> 81

Ser ser Ser Tyr Tyr Trp Gly1 5 <210> 82 <211> 16

<212> PRT <213> Human <400> 82

Gly lie Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys 5er15 10 15 <210> 83 <211> 13

<212> PRT <213> Human <400> 83

Gin Arg Gly His ser Ser Gly Trp Trp Tyr Phe Asp Leu15 10 <210> 84 <211> 11

<212> PRT <213> Human <400> 84

Arg Ala ser Gin Gly lie ser ser Tyr Leu Ala15 10 100 191718 <210> 85 ABXAZ 004VPC (2).TXT <211> 7 <212> PRT <213> Human <400> 85 Ala Ala Ser ser Leu Gin ser 1 5 <210> 86 <211> 9

<212> PRT <213> Human <400> 86

Gin Gin Ala Asn Asn Phe Pro Phe Thr1 5 <210> 87 . <211> 7

<212> PRT <213> Human <400> 87

Ser Ser Ser Asn Tyr Trp Gly1 5 <210> 88 <211> 16

<212> PRT <213> Human <400> 88

Gly lie Tyr Tyr Ser Gly ser Thr Tyr Tyr Asn Pro Ser Leu Arg Ser15 10 15 <210> 89 <211> 13

<212> PRT <213> Human <400> 89

Gin Arg Gly His Ser ser Gly Trp Trp Tyr Phe Asp Leu15 10 <210> 90 <211> 11

<212> PRT <213> Human <400> 90

Arg Ala Ser Arg Gly lie Ser Ser Trp Leu Ala 1 <210> 91 <211> 7 <212> PRT <213> Human <400> 91 101 191718

Thr Ala ser Ser Leu Gin.ser ABXAZ 004VPC (2).TXT 1 5 <210> 92<211> 9<212> PRT<213> Human <400> 92 Gin Gin Ala Asn ser Phe Pro Phe Thr 1 <210> 93<211> 25<212> PRT<213> Human 5 λ <400> 93

Gin val Gin Leu val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ala15 10 15

Ser val Lys Val Ser Cys Lys Ala Ser20 25 <210> 94 <211> 10

<212> PRT <213> Human <400> 94

Gly Tyr Thr Phe Thr ser Tyr Asp lie Asn15 10 <210> 95<211> 25<212> PRT<213> Human<400> 95

Gin Leu Gl n Leu Gl n Glu Ser Gly Pro Gly Leu val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr val Ser 20 25 <210> 96 <211> 12

<212> PRT <213> Human <400> 96

Gly Gly ser lie Arg Ser Ser Ser Tyr Tyr Trp Gly15 10 <210> 97<211> 25<212> PRT<213> Human <400> 97

Gin Leu Gl n Leu Gin Glu Ser Gly pro Gly Leu val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser 20 25 102 191718

ABXAZ 004VPC C2).TXT <210> 98 <211> 12

<212> PRT <213> Human <400> 98

Gly Gly Ser lie Asn Ser Ser Ser Asn Tyr Trp Gly15 10 <210> 99<211> 25<212> PRT<213> Human<400> 99

Gin val Gin Leu Gl n Glu Ser Gly Pro Gly Leu Val Lys Pro ser Glu 1 5 10 15 Thr Leu Ser Leu Thr cys Thr Val ser 20 25 <210> 100 <211> 10

<212> PRT <213> Human <400> 100

Gly Gly Ser lie Ser ser Tyr Tyr Trp ser15 10 <210> 101 <211> 25

<212> PRT <213> Human <400> 101

Gin val Gin Leu Gin Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu15 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser20 25 <210> 102 <211> 10

<212> PRT <213> Human <400> 102

Gly Gly Ser lie Ser Ser Tyr Tyr Trp Ser15 10 103 crairan ™*a , DTixan rw:n οτα mia^o pnvn irn πτ -|aoa,ρηη ηχΰπ laoana ma’na no^maa np’-ioo .zruwan rwao mpnan 7πίϊ: oxnmοιηπη 7v

<img img-format="tif" img-content="drawing" file="IL191718AD00021.tif" id="idf0001" />

.(moia nannn) erosion rwa