METHODS FOR TREATMENT OF POLYPOSIS

Abstract

The present invention relates to method for treating medical disorders mediated by mutations in the APC gene by administering an IGF1R inhibitor. Such disorders include, for example, familial adenomatous polyposis (FAP).

Description

[0001] This application is a divisional of U.S. patent application Ser. No. 11/834,303, filed Aug. 6, 2007; which claims the benefit of U.S. provisional patent application No. 60/835,941, filed Aug. 7, 2006; each of which is herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to methods for treating diseases mediated by the APC gene, such as familial adenomatous polyposis.

BACKGROUND OF THE INVENTION

[0003] Gardner Syndrome, Turcot Syndrome, familial adenomatous polyposis (FAP) and attenuated FAP are diseases that have been liked to the APC gene. APC is located on the long arm of chromosome 5.

[0004] Patients with FAP typically develop hundreds to thousands of colon polyps, usually starting in their teens. Essentially all subjects with FAP will develop colorectal cancer from the colon polyps usually by age 40. Patients with FAP often must have the colon, and sometimes the rectum, removed to prevent colon cancer. A less severe variant of FAP is attenuated FAP.

[0005] Gardner syndrome, a variant of FAP, is disease typically characterized by gastrointestinal polyps, multiple osteomas, and skin and soft tissue tumors. Cutaneous findings typically include epidermoid cysts, desmoid tumors, and other benign tumors. Polyps have essentially a 100% risk of undergoing malignant transformation.

[0006] Turcot Syndrome is a genetic disease typically characterized by polyps in the colon in addition to tumors in the brain. The polyps in the colon tend to become malignant. The brain tumors are also malignant. There are sometimes also skin abnormalities including cafe-au-lait (coffee-with-milk) spots, multiple lipomas (fatty tumors), and multiple scalp basal cell carcinoma (skin cancers of the scalp). Some consider Turcot syndrome to be a variant of FAP.

[0007] A study in mice demonstrated a connection between FAP and insulin-like growth factor II (IGF-II) (Hassan et al., Cancer Res. 60:1070-1076 (2000)). In this study, it was demonstrated that mutation of the IGF-II gene attenuated the expression of the adenomatous phenotype of mice with a mutation in the APC gene.

[0008] Presently, there are limited treatment options for patients suffering from FAP and other APC-mediated medical disorders. One of the first medications that was found to shrink colon polyps in patients with FAP was an anti-inflammatory medicine called sulindac. Sulindac is a commonly used nonsteroidal anti-inflammatory drug (NSAID). Many patients, however, cannot tolerate sulindac (e.g., due to gastrointestinal toxicity) and must discontinue taking it. An arthritis medication called celecoxib was approved by the FDA for the treatment of colon polyps in patients with FAP. Celecoxib, however, is a COX-2 inhibitor, and such medications have been linked recently to heart attacks (see e.g., Mukherjee et al., JAMA. 286:954-959 (2001)). Another routine treatment option is the drastic surgical measure of colectomy.

[0009] There clearly remains a need in the art for effective treatments of FAP and other APC-related medical disorders.

SUMMARY OF THE INVENTION

[0010] The present invention addresses the need in the art for additional treatments of FAP and other APC-related medical disorders by providing the methods and compositions of the invention as set forth herein.

[0011] The present invention comprises a method for treating or preventing an APC-mediated medical disorder in a subject, by administering, to the subject (e.g., a human), a therapeutically effective amount of an IGF1R inhibitor or a pharmaceutical composition thereof. In an embodiment of the invention, the disorder is a member selected from the group consisting of familial adenomatous polyposis (FAP), Gardner syndrome, Turcot syndrome and attenuated familial adenomatous polyposis. In an embodiment of the invention, the inhibitor is one or more members selected from the group consisting of an isolated antibody or antigen-binding fragment thereof that binds specifically to IGF1R (e.g., a monoclonal antibody, a polyclonal antibody, an anti-idiotypic antibody, a chimeric antibody, a bispecific antibody, a humanized antibody, a fully human antibody, a recombinant antibody, a Fab, a F(ab)2, an Fv, a single chain antibody, a dsFv and a linear antibody),

##STR00001##

[0012] In an embodiment of the invention, the antibody or antigen-binding fragment thereof comprises one or more CDRs from a light chain variable region comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, or 10 and/or one or more CDRs from a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 or 12. In an embodiment of the invention, the antibody is an isolated antibody or antigen binding fragment thereof that binds to IGF1R comprising a light chain immunoglobulin amino acid sequence which comprises CDR-L1 comprising amino acids RASQSIGSSLH (SEQ ID NO: 106), CDR-L2 comprising amino acids YASQSLS (SEQ ID NO: 107) and CDR-L3 comprising amino acids HQSSRLPHT (SEQ ID NO: 108); and/or a heavy chain immunoglobulin amino acid sequence which comprises CDR-H1 comprising amino acids SFAMH (SEQ ID NO: 109) or GFTFSSFAMH (SEQ ID NO: 110), CDR-H2 comprising amino acids VIDTRGATYYADSVKG (SEQ ID NO: 111) and CDR-H3 comprising amino acids LGNFYYGMDV (SEQ ID NO: 112). In an embodiment of the invention, the antibody is an isolated antibody comprising a light chain variable region comprising amino acids 20-128 of SEQ ID NO: 2, 4, 6 or 8 and/or a heavy chain variable region comprising amino acids 20-137 of SEQ ID NO: 10 or 12. In an embodiment of the invention, the antibody or fragment comprises a light chain immunoglobulin variable region linked to a kappa light chain constant region. In an embodiment of the invention, the antibody or fragment comprises a heavy chain immunoglobulin variable region linked to a gamma 1 heavy chain constant region. In an embodiment of the invention, the antibody or fragment comprises a mature heavy chain immunoglobulin encoded by a polynucleotide in plasmid 15H12/19D12 HCA (γ4) which is obtainable from a cell line deposited at the American Type Culture Collection (ATCC) under number PTA-5214; and/or a mature light chain immunoglobulin encoded by a polynucleotide in plasmid 15H12/19D12 LCF (κ) which is obtainable from a cell line deposited at the American Type Culture Collection (ATCC) under number PTA-5220. In an embodiment of the invention, the IGF1R inhibitor is administered to the subject in association with one or more therapeutic procedures, diagnostic procedures or additional therapeutic agents (e.g., a non-steroidal anti-inflammatory agent) e.g., in a single or separate compositions. In an embodiment of the invention, the additional therapeutic agent is one or members selected from the group consisting of any nonsteroidal anti-inflammatory agent (NSAID), any selective COX-2 inhibitor, piroxicam, sulindac, aptosyn (sulindac sulfone), indomethacin, rofecoxib, celecoxib or aspirin, a dietary calcium supplement, a retinoid, a carotenoid, ascorbic acid, α-tocopherol, selenium, folate and methionine. In an embodiment of the invention, the additional therapeutic agent is in a pharmaceutical composition in association with a pharmaceutically acceptable carrier. In an embodiment of the invention, the IGF1R inhibitor is administered to the subject by a parenteral route.

[0013] The present invention also provides a composition comprising one or more IGF1R inhibitors (e.g., anti-IGF1R antibodies, e.g., as set forth herein) in association with one or more agents that are suitable for treating or preventing any APC-mediated medical disorder such as FAP including any non-steroidal anti-inflammatory agent, any selective COX-2 inhibitor, piroxicam, sulindac, aptosyn (sulindac sulfone), indomethacin, rofecoxib, celecoxib, aspirin, a dietary calcium supplement, a retinoid, a carotenoid, ascorbic acid, α-tocopherol, selenium, folate and methionine) or a pharmaceutical composition thereof (i.e., comprising a pharmaceutically acceptable carrier).

DETAILED DESCRIPTION OF THE INVENTION

[0014] The present invention provides methods for treating or preventing any APC-mediated medical disorder. APC-mediated medical disorders include, for example, FAP, Gardner syndrome, Turcot Syndrome and attenuated FAP. Such disorders are treated by administering an IGF1R inhibitor to the subject optionally in association with a further therapeutic agent (e.g., an anti-inflammatory drug such as an NSAID) or in association with a therapeutic or diagnostic procedure, such as X-ray, colonoscopy or surgery.

[0015] The terms "IGF1R", "IGFR1", "Insulin-like Growth Factor Receptor-I" and "Insulin-like Growth Factor Receptor, type I" are well known in the art. Although IGF1R may be from any organism, in an embodiment of the invention it is from an animal, such as a mammal (e.g., mouse, rat, rabbit, sheep or dog) including a human. The nucleotide and amino acid sequence of a typical human IGF1R precursor has the Genbank Accession No. X04434 or NM_--000875. Cleavage of the precursor (e.g., between amino acids 710 and 711) produces an α-subunit and a β-subunit which associate to form a mature receptor.

[0016] As stated above, the present invention comprises methods for treating or preventing any APC-mediated disease or medical disorder. Such diseases or disorders include, for example, familial adenomatous polyposis (FAP; also known as familial polyposis coli), Gardner syndrome (see e.g., Gardner et al. Am. J. Hum. Genet. 5:139-47 (1953)), Turcot syndrome and attenuated FAP.

[0017] In subjects suffering from familial adenomatous polyposis, colorectal adenomatous polyps typically begin to appear at an average age of 16 years (range 7-36 years). In general, by age 35 years, 95% of individuals have polyps. Once they appear, the polyps generally rapidly increase in number; when colonic expression is fully developed, hundreds to thousands of colonic adenomatous polyps are typically observed. Without colectomy, colon cancer is essentially inevitable. The average age of colon cancer diagnosis in untreated individuals is 39 years (range 34-43 years). Other symptoms that can appear in conjunction with FAP are gastric polyps, adenomatous polyps of the small bowel, and extraintestinal manifestations including osteomas, dental abnormalities (e.g., unerupted teeth, congenital absence of one or more teeth, supernumerary teeth, dentigerous cysts, and odontomas), congenital hypertrophy of the retinal pigment epithelium (CHRPE), benign cutaneous lesions, desmoid tumors, adrenal masses and extracolonic cancers (e.g., small bowel, stomach, pancreas, thyroid, central nervous system, liver, bile ducts and adrenal glands).

[0018] Individuals with Gardner syndrome (GS) typically suffer from the colonic adenomatous polyposis of classic FAP with osteomas, and soft tissue tumors. Osteomas are typically found on the mandible and skull, although any bone of the body may be involved. Epidermoid cysts occur on any cutaneous surface and are mainly of cosmetic concern, as they do not appear to have malignant potential.

[0019] Individual suffering from Turcot syndrome typically exhibit colonic adenomatous polyposis of classic FAP and CNS tumors such as medulloblastoma. The risk of CNS tumors is substantially increased in persons with FAP generally, although the absolute risk is only approximately 1° A.

[0020] Attenuated FAP (AFAP) is typically characterized by a significant risk for colon cancer, but fewer colonic polyps (average of 30) than classic FAP. Polyps tend to be found more proximally in the colon than in classic FAP. The average age of colon cancer diagnosis in individuals with AFAP is age 50-55 years--10-15 years later than in those with classic FAP, but earlier than that seen in individuals with sporadically occurring colon cancer. Upper gastrointestinal polyps and cancers may be seen in individuals with AFAP and, although the extraintestinal manifestations of FAP may be present, CHRPE lesions and desmoid tumors are rare.

IGF1R Inhibitors

[0021] The present invention comprises methods for treating or preventing any APC-mediated medical disorder such as FAP, in a subject, by administering one or more IGF1R inhibitors to the subject.

[0022] The term "IGF1R inhibitor" includes any substance that decreases the expression, ligand binding, kinase activity or any other biological or biochemical activity (e.g., promotion of cellular growth) of IGF1R that will elicit a biological or medical response of a tissue, system or subject that is being sought by the administrator (such as a researcher, doctor or veterinarian) which includes any measurable alleviation of the signs, symptoms and/or clinical indicia of an APC-mediated medical disorder (e.g., FAP). An IGF1R inhibitor can be a small organic or inorganic molecule, or a large molecule such as a polypeptide, an antibody, an antigen-binding fragment of an antibody, a polymer or a nucleic acid.

[0023] In an embodiment of the invention, an IGF1R inhibitor that is administered to a patient in a method according to the invention is any isolated anti-insulin-like growth factor receptor-1 (IGF1R) antibody or antigen-binding fragment thereof (e.g., monoclonal antibodies (e.g., fully human monoclonal antibodies), polyclonal antibodies, bispecific antibodies, Fab antibody fragments, F(ab)₂ antibody fragments, Fv antibody fragments (e.g., VH or VL), linear antibodies, single chain Fv antibody fragments, dsFv antibody fragments, humanized antibodies, chimeric antibodies or anti-idiotypic antibodies) such as any of those disclosed in any of Burtrum et. al Cancer Research 63:8912-8921 (2003); in French Patent Applications FR2834990, FR2834991 and FR2834900 and in PCT Application Publication Nos. WO 03/100008; WO 03/59951; WO 04/71529; WO 03/106621; WO 04/83248; WO 04/87756 and WO 02/53596.

[0024] In an embodiment of the invention, an IGF1R inhibitor that is administered to a patient in a method according to the invention is an isolated anti-insulin-like growth factor receptor-1 (IGF1R) antibody or antigen-binding fragment thereof comprising a mature or unprocessed 19D12/15H12 Light Chain-C, D, E or F and/or a mature 19D12/15H12 heavy chain-A or B. In an embodiment of the invention, an IGF1R inhibitor that is administered to a patient in a method according to the invention is an isolated antibody that specifically binds to IGF1R that comprises one or more complementarity determining regions (CDRs) of 19D12/15H12 Light Chain-C, D, E or F and/or 19D12/15H12 heavy chain-A or B (e.g., all 3 light chain CDRs and all 3 heavy chain CDRs).

[0025] The amino acid and nucleotide sequences of the 19D12/15H12 antibody chains are shown below. Dotted, underscored type indicates the signal peptide. Solid underscored type indicates the CDRs. Plain type indicates the framework regions. Mature fragments lack the signal peptide. Such sequences are also disclosed in published U.S. patent application no. US 2004/0018191 and in published international patent application no. WO 03/100008; each of which is herein incorporated by reference in its entirety.

TABLE-US-00001 Modified 19D12/15H12 Light Chain-C (SEQ ID NO: 1) ##STR00002## GGC GAG AGA GTC ACC ATC ACC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG TCT CCA AAG CTT CTC ATC AAG TAT GCA TCC CAG TCC CTC TCA GGG GTC CCC TCG AGG TTC AGT GGC AGT GGA TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGC CTC GAG GCT GAA GAT GCT GCA GCG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA GGG ACC AAG GTG GAG ATC AAA CGT ACG (SEQ ID NO: 2) ##STR00003## G E R V T I T C R A S Q S I G S S L H W Y Q Q K P G Q S P K L L I K Y A S Q S L S G V P S R F S G S G S G T D F T L T I S S L E A E D A A A Y Y C H Q S S R L P H T F G Q G T K V E I K R T Modified 19D12/15H12 Light Chain-D (SEQ ID NO: 3) ##STR00004## GGC GAG AGA GTC ACC ATC ACC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG TCT CCA AAG CTT CTC ATC AAG TAT GCA TCC CAG TCC CTC TCA GGG GTC CCC TCG AGG TTC AGT GGC AGT GGA TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGC CTC GAG GCT GAA GAT TTC GCA GTG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA GGG ACC AAG GTG GAG ATC AAA CGT ACG (SEQ ID NO: 4) ##STR00005## G E R V T I T C R A S Q S I G S S L H W Y Q Q K P G Q S P K L L I K Y A S Q S L S G V P S R F S G S G S G T D F T L T I S S L E A E D F A V Y Y C H Q S S R L P H T F G Q G T K V E I K R T Modified 19D12/15H12 Light Chain-E (SEQ ID NO: 5) ##STR00006## GGC GAG AGA GCC ACC CTC TCC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG GCT CCA AGG CTT CTC ATC AAG TAT GCA TCC CAG TCC CTC TCA GGG ATC CCC GAT AGG TTC AGT GGC AGT GGA TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGA CTC GAG CCT GAA GAT GCT GCA GCG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA GGG ACC AAG GTG GAG ATC AAA CGT ACA (SEQ ID NO: 6) ##STR00007## G E R A T L S C R A S Q S I G S S L H W Y Q Q K P G Q A P R L L I K Y A S Q S L S G I P D R F S G S G S G T D F T L T I S R L E P E D A A A Y Y C H Q S S R L P H T F G Q G T K V E I K R T 19D12/15H12 Light Chain-F (LCF; SEQ ID NO: 7) ##STR00008## GGC GAG AGA GCC ACC CTC TCC TGC CGG GCC AGT CAG AGC ATT GGT AGT AGC TTA CAC TGG TAC CAG CAG AAA CCA GGT CAG GCT CCA AGG CTT CTC ATC AAG TAT GCA TCC CAG TCC CTC TCA GGG ATC CCC GAT AGG TTC AGT GGC AGT GGA TCT GGG ACA GAT TTC ACC CTC ACC ATC AGT AGA CTG GAG CCT GAA GAT TTC GCA GTG TAT TAC TGT CAT CAG AGT AGT CGT TTA CCT CAC ACT TTC GGC CAA GGG ACC AAG GTG GAG ATC AAA CGT ACA (SEQ ID NO: 8) ##STR00009## G E R A T L S C R A S Q S I G S S L H W Y Q Q K P G Q A P R L L I K Y A S Q S L S G I P D R F S G S G S G T D F T L T I S R L E P E D F A V Y Y C H Q S S R L P H T F G Q G T K V E I K R T 19D12/15H12 heavy chain-A (HCA; SEQ ID NO: 9) ##STR00010## GGG TCC CTG AGA CTC TCC TGT GCA GCC TCT GGA TTC ACC TTC AGT AGC TTT GCT ATG CAC TGG GTT CGC CAG GCT CCA GGA AAA GGT CTG GAG TGG ATA TCA GTT ATT GAT ACT CGT GGT GCC ACA TAC TAT GCA GAC TCC GTG AAG GGC CGA TTC ACC ATC TCC AGA GAC AAT GCC AAG AAC TCC TTG TAT CTT CAA ATG AAC AGC CTG AGA GCC GAG GAC ACT GCT GTG TAT TAC TGT GCA AGA CTG GGG AAC TTC TAC TAC GGT ATG GAC GTC TGG GGC CAA GGG ACC ACG GTC ACC GTC TCC TCA (SEQ ID NO: 10) ##STR00011## Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Modified 19D12/15H12 heavy chain-B (SEQ ID NO: 11) ##STR00012## GGG TCC CTG AGA CTC TCC TGT GCA GCC TCT GGA TTC ACC TTC AGT AGC TTT GCT ATG CAC TGG GTT CGC CAG GCT CCA GGA AAA GGT CTG GAG TGG ATA TCA GTT ATT GAT ACT CGT GGT GCC ACA TAC TAT GCA GAC TCC GTG AAG GGC CGA TTC ACC ATC TCC AGA GAC AAT GCC AAG AAC TCC TTG TAT CTT CAA ATG AAC AGC CTG AGA GCC GAG GAC ACT GCT GTG TAT TAC TGT GCA AGA CTG GGG AAC TTC TAC TAC GGT ATG GAC GTC TGG GGC CAA GGG ACC ACG GTC ACC GTC TCC TCA (SEQ ID NO: 12) ##STR00013## Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

[0026] Cell lines comprising plasmids comprising a CMV promoter operably linked to the 15H12/19D12 LCC, LCD, LCE, LCF or to the 15H12/19D12 HCA or HCB have been deposited at the American Type Culture Collection (ATCC); 10801 University Boulevard; Manassas, Va. 20110-2209 on May 21, 2003. The deposit names and the ATCC accession numbers for the cell lines are set forth below:

[0027] (1) CMV promoter-15H12/19D12 HCA (γ4)--

[0028] Deposit name: "15H12/19D12 HCA (γ4)"

[0029] ATCC accession No.: PTA-5214

[0030] (2) CMV promoter-15H12/19D12 HCB (γ4)--

[0031] Deposit name: "15H12/19D12 HCB (γ4)"

[0032] ATCC accession No.: PTA-5215

[0033] (3) CMV promoter-15H12/19D12 HCA (γ1)--

[0034] Deposit name: "15H12/19D12 HCA (γ1)";

[0035] ATCC accession No.: PTA-5216

[0036] (4) CMV promoter-15H12/19D12 LCC (κ)--

[0037] Deposit name: "15H12/19D12 LCC (κ)";

[0038] ATCC accession No.: PTA-5217

[0039] (5) CMV promoter-15H12/19D12 LCD (κ)--

[0040] Deposit name: "15H12/19D12 LCD (κ)";

[0041] ATCC accession No.: PTA-5218

[0042] (6) CMV promoter-15H12/19D12 LCE (κ)--

[0043] Deposit name: "15H12/19D12 LCE (κ)";

[0044] ATCC accession No.: PTA-5219

[0045] (7) CMV promoter-15H12/19D12 LCF (κ)--

[0046] Deposit name: "15H12/19D12 LCF (κ)";

[0047] ATCC accession No.: PTA-5220

[0048] All restrictions on access to the plasmids deposited in ATCC will be removed upon grant of a patent. In an embodiment of the present invention, an anti-IGF1R antibody or antigen-binding fragment thereof of the invention comprises any of the CDRs or Ig heavy or light chains or variable regions thereof (mature or unprocessed) in any of PTA-5214-PTA-5220. In an embodiment of the invention, the antibody comprises a mature (lacking signal sequence) light chain encoded by the plasmid deposited under number PTA-5220 and a mature (lacking signal sequence) heavy chain encoded by the plasmid deposited under number PTA-5214 or PTA-5216.

[0049] In an embodiment of the invention, the anti-IGF1R antibody or antigen-binding fragment thereof comprises one or more (e.g., 3) CDRs taken from 19D12/15H12 LC-C, D, E or F and/or one or more (e.g., 3) CDRs taken from 19D12/15H12 HC-A or B or any other immunoglobulin set forth herein as identified by the Chothia and/or Kabat system for identifying immunoglobulin domains (Chothia et al., J. Mol. Biol. 186:651-663 (1985); Novotny and Haber, Proc. Natl. Acad. Sci. USA 82:4592-4596 (1985) or Kabat, E. A. et al., Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., (1987)).

[0050] In an embodiment, an antibody that binds "specifically" to human IGF1R binds with Kd of about 1.28×10^-10 M or less by Biacore measurement or with a Kd of about 2.05×10^-12 or less by KinExA measurement.

[0051] In an embodiment of the invention, an IGF1R inhibitor that can be administered to a patient in a method according to the invention comprises any light chain immunoglobulin and/or a heavy chain immunoglobulin as set forth in Published International Application No. WO 2002/53596 which is herein incorporated by reference in its entirety. For example, in an embodiment, the antibody comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 6, 10, 14, 18, 22, 47 and 51 as set forth in WO 2002/53596 and/or a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 8, 12, 16, 20, 24, 45 and 49 as set forth in WO 2002/53596.

[0052] In an embodiment of the invention, an IGF1R inhibitor that can be administered to a patient in a method according to the invention comprises any light chain immunoglobulin and/or a heavy chain immunoglobulin as set forth in Published International Application No. WO 2003/59951 which is herein incorporated by reference in its entirety. For example, in an embodiment, the antibody comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 54, 61 and 65 as set forth in WO 2003/59951 and/or a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 69, 75, 79 and 83 as set forth in WO 2003/59951.

[0053] In an embodiment of the invention, an IGF1R inhibitor that can be administered to a patient in a method according to the invention comprises any light chain immunoglobulin and/or a heavy chain immunoglobulin as set forth in Published International Application No. WO 2004/83248 which is herein incorporated by reference in its entirety. For example, in an embodiment, the antibody comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141 and 143 as set forth in WO 2004/83248 and/or a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140 and 142 as set forth in WO 2004/83248.

[0054] In an embodiment of the invention, an IGF1R inhibitor that can be administered to a patient in a method according to the invention comprises any light chain immunoglobulin and/or a heavy chain immunoglobulin as set forth in Published International Application No. WO 2003/106621 which is herein incorporated by reference in its entirety. For example, in an embodiment, the antibody comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 8-12, 58-69, 82-86, 90, 94, 96, 98, as set forth in WO 2003/106621 and/or a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 7, 13, 70-81, 87, 88, 92 as set forth in WO 2003/106621.

[0055] In an embodiment of the invention, an IGF1R inhibitor that can be administered to a patient in a method according to the invention comprises any light chain immunoglobulin and/or a heavy chain immunoglobulin as set forth in Published International Application No. WO 2004/87756 which is herein incorporated by reference in its entirety. For example, in an embodiment, the antibody comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 2 as set forth in WO 2004/87756 and/or a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1 as set forth in WO 2004/87756.

[0056] Furthermore, the scope of the present invention comprises any antibody or antibody fragment comprising one or more CDRs and/or framework regions of any of the light chain immunoglobulin or heavy chain immunoglobulins set forth in WO 2002/53596; WO 2003/59951; WO 2004/83248; WO 2003/106621 or WO 2004/87756 as identified by any of the methods set forth in Chothia et al., J. Mol. Biol. 186:651-663 (1985); Novotny and Haber, Proc. Natl. Acad. Sci. USA 82:4592-4596 (1985) or Kabat, E. A. et al., Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., (1987)).

[0057] In an embodiment of the invention, anti-IGF1R antibody is produced by a hybridoma that is deposited at the American Type Culture Collection under deposit no. PTA-2792, PTA-2788, PTA-2790, PTA-2791, PTA-2789 or PTA-2793.

[0058] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises an immunoglobulin heavy chain variable region comprising an amino acid sequence selected from the group consisting of:

TABLE-US-00002 (SEQ ID NO: 13) 1 grlgqawrsl rlscaasgft fsdyymswir qapgkglewv syisssgstr 51 dyadsvkgrf tisrdnakns lylqmnslra edtavyycvr dgvettfyyy 101 yygmdvwgqg ttvtvssast kgpsvfplap csrstsesta algclvkdyf 151 pepvtvswns galtsgvhtf psca (SEQ ID NO: 14) 1 vqllesgggl vqpggslrls ctasgftfss yamnwvrqap gkglewvsai 51 sgsggttfya dsvkgrftis rdnsrttlyl qmnslraedt avyycakdlg 101 wsdsyyyyyg mdvwgqgttv tvss (SEQ ID NO: 15) 1 gpglvkpset lsltctvsgg sisnyywswi rqpagkglew igriytsgsp 51 nynpslksrv tmsvdtsknq fslklnsvta adtavyycav tifgvviifd 101 ywgqgtlvtv ss (SEQ ID NO: 16) 1 evqllesggg lvqpggslrl scaasgftfs syamswvrqa pgkglewvsa 51 isgsggityy adsvkgrfti srdnskntly lqmnslraed tavyycakdl 101 gygdfyyyyy gmdvwgqgtt vtvss (SEQ ID NO: 17) 1 pglvkpsetl sltctvsggs issyywswir qppgkglewi gyiyysgstn 51 ynpslksrvt isvdtsknqf slklssvtaa dtavyycart ysssfyyygm 101 dvwgqgttvt vss (SEQ ID NO: 18) 1 evqllesggg lvqpggslrl scaasgftfs syamswvrqa pgkglewvsg 51 itgsggstyy adsvkgrfti srdnskntly lqmnslraed tavyycakdp 101 gttvimswfd pwgqgtlvtv ss

[0059] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises an immunoglobulin light chain variable region comprising an amino acid sequence selected from the group consisting of:

TABLE-US-00003 (SEQ ID NO: 19) 1 asvgdrvtft crasqdirrd lgwyqqkpgk apkrliyaas rlqsgvpsrf 51 sgsgsgteft ltisslqped fatyyclqhn nyprtfgqgt eveiirtvaa 101 psvfifppsd eqlksgtasv vcllnnfypr eakvqw (SEQ ID NO: 20) 1 diqmtqfpss lsasvgdrvt itcrasqgir ndlgwyqqkp gkapkrliya 51 asrlhrgvps rfsgsgsgte ftltisslqp edfatyyclq hnsypcsfgq 101 gtkleik (SEQ ID NO: 21) 1 sslsasvgdr vtftcrasqd irrdlgwyqq kpgkapkrli yaasrlqsgv 51 psrfsgsgsg teftltissl qpedfatyyc lqhnnyprtf gqgteveiir (SEQ ID NO: 22) 1 diqmtqspss lsasvgdrvt itcrasqgir sdlgwfqqkp gkapkrliya 51 asklhrgvps rfsgsgsgte ftltisrlqp edfatyyclq hnsypltfgg 101 gtkveik (SEQ ID NO: 23) 1 gdrvtitcra sqsistflnw yqqkpgkapk llihvasslq ggvpsrfsgs 51 gsgtdftlti sslqpedfat yycqqsynap ltfgggtkve ik (SEQ ID NO: 24) 1 ratlscrasq svrgrylawy qqkpgqaprl liygassrat gipdrfsgsg 51 sgtdftltis rlepedfavf ycqqygsspr tfgqgtkvei k

[0060] In an embodiment of the invention, the anti-IGF1R antibody comprises a light chain immunoglobulin, or a mature fragment thereof (i.e., lacking signal sequence), or variable region thereof, comprising the amino acid sequence of:

TABLE-US-00004 (SEQ ID NO: 25) 1 mdmrvpaqll gllllwfpga rcdiqmtqsp sslsasvgdr vtitcrasqg 51 irndlgwyqq kpgkapkrli yaasslqsgv psrfsgsgsg teftltissl 101 qpedfatyyc lqhnsypwtf gqgtkveikr tvaapsvfif ppsdeqlksg 151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst 201 ltlskadyek hkvyacevth qglsspvtks fnrgec; (SEQ ID NO: 26) 1 mdmrvpaqll gllllwfpga rcdiqmtqsp sslsasvgdr vtftcrasqd 51 irrdlgwyqq kpgkapkrli yaasrlqsgv psrfsgsgsg teftltissl 101 qpedfatyyc lqhnnyprtf gqgteveiir tvaapsvfif ppsdeqlksg 151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst 201 ltlskadyek hkvyacevth qglsspvtks fnrgec; (SEQ ID NO: 27) 1 mdmrvpaqll gllllwfpga rcdiqmtqsp sslsasvgdr vtiterasqg 51 irndlgwyqq kpgkapkrli yaasslqsgv psrfsgsgsg teftltissl 101 qpedfatyyc lqhnsypytf gqgtkleikr tvaapsvfif ppsdeqlksg 151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst 201 ltlskadyek hkvyacevth qglsspvtks fnrgec; or (SEQ ID NO: 28) 1 mdmrvpaqll gllllwfpga rcdiqmtqfp sslsasvgdr vtitcrasqg 51 irndlgwyqq kpgkapkrli yaasrlhrgv psrfsgsgsg teftltissl 101 qpedfatyyc lqhnsypcsf gqgtkleikr tvaapsvfif ppsdeqlksg 151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvteqds kdstyslsst 201 ltlskadyek hkvyacevth qglsspvtks fnrgec.

(SEQ ID NO: 28). In an embodiment of the invention, the signal sequence is amino acids 1-22 of SEQ ID NOs: 25-28. In an embodiment of the invention, the mature variable region is underscored.

[0061] In an embodiment of the invention, the anti-IGF1R antibody comprises a heavy chain immunoglobulin or a mature fragment thereof (i.e., lacking signal sequence), or a variable region thereof, comprising the amino acid sequence of:

TABLE-US-00005 (SEQ ID NO: 29) 1 mefglswvfl vaiikgvqcq vqlvesgggl vkpggslrls caasgftfsd 51 yymswirqap gkglewvsyi sssgstiyya dsvkgrftis rdnaknslyl 101 qmnslraedt avyycarvlr flewllyyyy yygmdvwgqg ttvtvssast 151 kgpsvfplap csrstsesta algclvkdyf pepvtvswns galtsgvhtf 201 pavlqssgly slssvvtvps snfgtqtytc nvdhkpsntk vdktverkcc 251 vecppcpapp vagpsvflfp pkpkdtlmis rtpevtcvvv dvshedpevq 301 fnwyvdgvev hnaktkpree qfnstfrvvs vltvvhqdwl ngkeykckvs 351 nkglpapiek tisktkgqpr epqvytlpps reemtknqvs ltclvkgfyp 401 sdiavewesn gqpennyktt ppmldsdgsf flyskltvdk srwqqgnvfs 451 csvmhealhn hytqkslsls pgk; (SEQ ID NO: 30) 1 mefglswvfl vaiikgvqcq aqlvesgggl vkpggslrls caasgftfsd 51 yymswirqap gkglewvsyi sssgstrdya dsvkgrftis rdnaknslyl 101 qmnslraedt avyycvrdgv ettfyyyyyg mdvwgqgttv tvssastkgp 151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav 201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec 251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw 301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg 351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi 401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv 451 mhealhnhyt qkslslspgk; (SEQ ID NO: 31) 1 mefglswlfl vailkgvqce vqllesgggl vqpggslrls caasgftfss 51 yamswvrqap gkglewvsai sgsggstyya dsvkgrftis rdnskntlyl 101 qmnslraedt avyycakgys sgwyyyyyyg mdvwgqgttv tvssastkgp 151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav 201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec 251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw 301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg 351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi 401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv 451 mhealhnhyt qkslslspgk; or (SEQ ID NO: 32) 1 mefglswlfl vailkgvqce vqllesgggl vqpggslrls ctasgftfss 51 yamnwvrqap gkglewvsai sgsggttfya dsvkgrftis rdnsrttlyl 101 qmnslraedt avyycakdlg wsdsyyyyyg mdvwgqgttv tvssastkgp 151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav 201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec 251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw 301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg 351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi 401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv 451 mhealhnhyt qkslslspgk.

In an embodiment of the invention, the signal sequence is amino acids 1-19 of SEQ ID NOs: 29-32. In an embodiment of the invention, the mature variable region is underscored.

[0062] In an embodiment of the invention, the anti-IGF1R antibody comprises a light chain variable region comprising the amino acid sequence of any of SEQ ID NOs: 19-24 paired with a heavy chain variable region comprising an amino acid sequence of any of SEQ ID NOs: 13-18, respectively. In an embodiment of the invention, the anti-IGF1R antibody comprises a mature light chain variable region comprising an amino acid sequence of any of SEQ ID NOs: 25 or 26 paired with a heavy chain variable region comprising an amino acid sequence of any of SEQ ID NOs: 29 or 30. In an embodiment of the invention, the anti-IGF1R antibody comprises a mature light chain variable region comprising an amino acid sequence of any of SEQ ID NOs: 27 or 28 paired with a heavy chain variable region comprising an amino acid sequence of any of SEQ ID NOs: 31 or 32.

[0063] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises an immunoglobulin heavy chain or mature fragment or variable region of 2.12.1 fx (SEQ ID NO: 33) (in an embodiment of the invention, the leader sequence is underscored):

TABLE-US-00006 1 mefglswvfl vaiikgvqcq vqlvesgggl vkpggslrls caasgftfsd 51 yymswirqap gkglewvsyi sssgstrdya dsvkgrftis rdnaknslyl 101 qmnslraedt avyycardgv ettfyyyyyg mdvwgqgttv tvssastkgp 151 svfplapcsr stsestaalg clvkdyfpep vtvswnsgal tsgvhtfpav 201 lqssglysls svvtvpssnf gtqtytcnvd hkpsntkvdk tverkccvec 251 ppcpappvag psvflfppkp kdtlmisrtp evtcvvvdvs hedpevqfnw 301 yvdgvevhna ktkpreeqfn stfrvvsvlt vvhqdwlngk eykckvsnkg 351 lpapiektis ktkgqprepq vytlppsree mtknqvsltc lvkgfypsdi 401 avewesngqp ennykttppm ldsdgsffly skltvdksrw qqgnvfscsv 451 mhealhnhyt qkslslspgk

[0064] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises mature immunoglobulin heavy chain variable region 2.12.1 fx (amino acids 20-144 or SEQ ID NO: 33; SEQ ID NO: 34):

TABLE-US-00007 q vqlvesgggl vkpggslrls caasgftfsd yymswirqap gkglewvsyi sssgstrdya dsvkgrftis rdnaknslyl qmnslraedt avyycardgv ettfyyyyyg mdvwgqgttv tvss

[0065] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises an immunoglobulin light chain or mature fragment or variable region 2.12.1 fx (SEQ ID NO: 35) (in an embodiment of the invention, the leader sequence is underscored):

TABLE-US-00008 1 mdmrvpaqll gllllwfpga rcdiqmtqsp sslsasvgdr vtitcrasqd 51 irrdlgwyqq kpgkapkrli yaasrlqsgv psrfsgsgsg teftltissl 101 qpedfatyyc lqhnnyprtf gqgtkveikr tvaapsvfif ppsdeqlksg 151 tasvvcllnn fypreakvqw kvdnalqsgn sqesvtegds kdstyslsst 201 ltlskadyek hkvyacevth qglsspvtks fnrgec

[0066] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises mature immunoglobulin light chain variable region 2.12.1 fx (amino acids 23-130 of SEQ ID NO: 35; SEQ ID NO: 36):

TABLE-US-00009 diqmtqsp sslsasvgdr vtitcrasqd irrdlgwyqq kpgkapkrli yaasrlqsgv psrfsgsgsg teftltissl qpedfatyyc lqhnnyprtf gqgtkveikr

[0067] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises a humanized 7C10 immunoglobulin light chain variable region; version 1 (SEQ ID NO: 37):

TABLE-US-00010 1 dvvmtqspls lpvtpgepas iscrssqsiv hsngntylqw ylqkpgqspq 51 lliykvsnrl ygvpdrfsgs gsgtdftlki srveaedvgv yycfqgshvp 101 wtfgqgtkve ik

[0068] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises humanized 7C10 immunoglobulin light chain variable region; version 2 (SEQ ID NO: 38):

TABLE-US-00011 1 divmtqspls 1pvtpgepas iscrssqsiv hsngntylqw ylqkpgqspq 51 lliykvsnrl ygvpdrfsgs gsgtdftlki srveaedvgv yycfqgshvp 101 wtfgqgtkve ik

[0069] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises a humanized 7C10 immunoglobulin heavy chain variable region; version 1 (SEQ ID NO: 39):

TABLE-US-00012 1 qvglgesgpg lvkpsetlsl tctvsgysit ggylwnwirq ppgkglewmg 51 yisydgtnny kpslkdriti srdtsknqfs lklssvtaad tavyycaryg 101 rvffdywgqg tlvtvss

[0070] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises the humanized 7C10 immunoglobulin heavy chain variable region; version 2 (SEQ ID NO: 40):

TABLE-US-00013 1 qvqlqesgpg lvkpsetlsl tctvsgysit ggylwnwirq ppgkglewig 51 yisydgtnny kpslkdrvti srdtsknqfs lklssvtaad tavyycaryg 101 rvffdywgqg tlvtvss

[0071] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises the humanized 7C10 immunoglobulin heavy chain variable region; version 3 (SEQ ID NO: 41):

TABLE-US-00014 1 qvqlqesgpg lvkpsetlsl tctvsgysis ggylwnwirq ppgkglewig 51 yisydgtnny kpslkdrvti svdtsknqfs lklssvtaad tavyycaryg 101 rvffdywgqg tivtvss

[0072] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises A12 immunoglobulin heavy chain variable region (SEQ ID NO: 42):

TABLE-US-00015 1 evqlvqsgae vkkpgssvkv sckasggtfs syaiswvrqa pgqglewmgg 51 iipifgtany aqkfqgrvti tadkststay melsslrsed tavyycarap 101 lrflewstqd hyyyyymdvw gkgttvtvss

[0073] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises A12 immunoglobulin light chain variable region (SEQ ID NO: 43):

TABLE-US-00016 1 sseltqdpav svalgqtvri tcqgdslrsy yaswyqqkpg qapvlviygk 51 nnrpsgipdr fsgsssgnta sltitgaqae deadyycnsr dnsdnrlifg 101 ggtkltvls

or

(SEQ ID NO: 105):

TABLE-US-00017

[0074] 1 sseltqdpav svalgqtvri tcqgdslrsy yatwyqqkpg qapilviyge 51 nkrpsgipdr fsgsssgnta sltitgaqae deadyycksr dgsgqhlvfg 101 ggtkltvlg

[0075] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises 1A immunoglobulin heavy chain variable region (SEQ ID NO: 44):

TABLE-US-00018 1 evqlvqsggg lvhpggslrl scagsgftfr nyamywvrqa pgkglewvsa 51 igsgggtyya dsvkgrftis rdnaknslyl qmnslraedm avyycarapn 101 wgsdafdiwg qgtmvtvss

;optionally including one or more of the following mutations: R30, S30, N31, S31, Y94, H94, D104, E104.

[0076] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises 1A immunoglobulin light chain variable region (SEQ ID NO: 45):

TABLE-US-00019 1 diqmtqspss lsasvgdrvt itcrasqgis swlawyqqkp ekapksliya 51 asslqsgvps rfsgsgsgtd ftltisslqp edfatyycqq ynsypptfgp 101 gtkvdik

[0077] ;optionally including one or more of the following mutations: P96, I96, P100, Q100, R103, K103, V104, L104, D105, E105

[0078] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises single chain antibody (fv) 8A1 (SEQ ID NO: 46):

TABLE-US-00020 1 evqlvqsgae vkkpgeslti sckgpgynff nywigwvrqm pgkglewmgi 51 iyptdsdtry spsfqgqvti svdksistay lqwsslkasd tamyycarsi 101 rycpggrcys gyygmdvwgq gtmvtvssgg ggsggggsgg ggsseltqdp 151 avsvalgqtv ritcqgdslr syyaswyqqk pgqapvlviy gknnrpsgip 201 drfsgsssgn tasltitgaq aedeadyycn srdssgnhvv fgggtkltvl 251 g

[0079] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises single chain antibody (fv) 9A2 (SEQ ID NO: 47):

TABLE-US-00021 1 qvqlvqsgae vrkpgasvkv scktsgytfr nydinwvrqa pgqglewmgr 51 isghygntdh aqkfqgrftm tkdtststay melrsltfdd tavyycarsq 101 wnvdywgrgt lvtvssgggg sggggsgggg salnfmltqp hsysespgkt 151 vtisctrssg siasnyvqwy qqrpgssptt vifednrrps gvpdrfsgsi 201 dtssnsaslt isglktedea dyycqsfdst nlvvfgggtk vtvlg

[0080] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises single chain antibody (fv) 11A4 (SEQ ID NO: 48):

TABLE-US-00022 1 evqllesggg lvqpggslrl scaasgftfs syamswvrqa pgkglewvsa 51 isgsggstyy adsvkgrfti srdnskntly lqmnslraed tavyycassp 101 yssrwysfdp wgqgtmvtvs sggggsgggg sggggsalsy eltqppsvsv 151 spgqtatitc sgddlgnkyv swyqqkpgqs pvlviyqdtk rpsgiperfs 201 gsnsgniatl tisgtqavde adyycqvwdt gtvvfgggtk ltvlg

[0081] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises single chain antibody (fv) 7A4 (SEQ ID NO: 49):

TABLE-US-00023 1 evqlvqsgae vkkpgeslti sckgsgynff nywigwvrqm pgkdlewmgi 51 iyptdsdtry spsfqgqvti svdksistay lqwsslkasd tamyycarsi 101 rycpggrcys gyygmdvwgq gtmvtvssgg gssggggsgg ggsseltqdp 151 avsvalgqtv ritcrgdslr nyyaswyqqk pgqapvlviy gknnrpsgip 201 drfsgsssgn tasltitgaq aedeadyycn srdssgnhmv fgggtkltvl 251 g

[0082] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises single chain antibody (fv) 11A1 (SEQ ID NO: 50):

TABLE-US-00024 1 evqlvesggg vvqpgrslrl scaasgftfs dfamhwvrgi pgkglewlsg 51 lrhdgstayy agsvkgrfti srdnsrntvy lqmnslraed tatyycvtgs 101 gssgphafpv wgkgtlvtvs sggggsgggg sggggsalsy vltqppsasg 151 tpgqrvtisc sgsnsnigty tvnwfqqlpg tapklliysn nqrpsgvpdr 201 fsgsksgtsa slaisglqse deadyycaaw ddslngpvfg ggtkvtvlg

[0083] In an embodiment of the invention, an anti-IGF1R antibody of the invention comprises single chain antibody (fv) 7A6 (SEQ ID NO: 51)

TABLE-US-00025 1 evqlvqsgae vkkpgeslti sckgsgynff nywigwvrqm pgkglewmgi 51 iyptdsdtry spsfqgqvti svdksistay lqwsslkasd tamyycarsi 101 rycpggrcys gyygmdvwgq gtlvtvssgg ggsggggsgg ggsseltqdp 151 avsvalgqtv ritcqgdslr syytnwfqqk pgqapllvvy aknkrpsgip 201 drfsgsssgn tasltitgaq aedeadyycn srdssgnhvv fgggtkltvl 251 g

[0084] In an embodiment of the invention, an anti-IGF1R antibody or an antigen-binding fragment thereof (e.g., a heavy chain or light chain immunoglobulin) of the invention comprises one or more complementarity determining regions (CDR) selected from the group consisting of:

TABLE-US-00026 (SEQ ID NO: 52) sywmh; (SEQ ID NO: 53) einpsngrtnynekfkr; (SEQ ID NO: 54) grpdyygsskwyfdv; (SEQ ID NO: 55) rssqsivhsnvntyle; (SEQ ID NO: 56) kvsnrfs; and (SEQ ID NO: 57) fqgshvppt.

[0085] In an embodiment of the invention, an anti-IGF1R antibody or an antigen-binding fragment thereof of the invention comprises a heavy chain immunoglobulin variable region selected from the group consisting of:

TABLE-US-00027 (SEQ ID NO: 58) 1 qvqlvqsgae vvkpgasvkl sckasgytft sywmhwvkqr pgqglewige 51 inpsngrtny nqkfqgkatl tvdkssstay mqlssltsed savyyfargr 101 pdyygsskwy fdvwgqgttv tvs; (SEQ ID NO: 59) 1 qvqfqqsgae lvkpgasvkl sckasgytft sylmhwikqr pgrglewigr 51 idpnnvvtkf nekfkskatl tvdkpsstay melssltsed savyycarya 101 ycrpmdywgq gttvtvss; (SEQ ID NO: 60) 1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgqglewige 51 inpsngrtny nekfkrkatl tvdkssstay mqlssltsed savyyfargr 101 pdyygsskwy fdvwgagttv tvs; (SEQ ID NO: 61) 1 qvqlqqsgae lmkpgasvki sckatgytfs sfwiewvkqr pghglewige 51 ilpgsggthy nekfkgkatf tadkssntay mqlssltsed savyycargh 101 syyfydgdyw gqgtsvtvss; (SEQ ID NO: 62) 1 qvqlqqpgsv lvrpgasvkl sckasgytft sswihwakqr pgqglewige 51 ihpnsgntny nekfkgkatl tvdtssstay vdlssltsed savyycarwr 101 ygspyyfdyw gqgttltvss; (SEQ ID NO: 63) 1 qvglqqpgae lvkpgasvkl sckasgytft sywmhwvkqr pgrglewigr 51 idpnsggtky nekfkskatl tvdkpsstay mqlssltsed savyycaryd 101 yygssyfdyw gqgttltvss; (SEQ ID NO: 64) 1 qvqlvqsgae vvkpgasvkl sckasgytft sywmhwvkqr pgqglewige 51 inpsngrtny nqkfqgkatl tvdkssstay mqlssltsed savyyfargr 101 pdyygsskwy fdvwgqgttv tvs; (SEQ ID NO: 65) 1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgqglewige 51 inpsngrtny nekfkrkatl tvdkssstay mqlssltsed savyyfargr 101 pdyygsskwy fdvwgagttv tvss; (SEQ ID NO: 66) 1 qvqlvqsgae vvkpgasvkl sckasgytft sywmhwvkqr pgqglewige 51 inpsngrtny nqkfqgkatl tvdkssstay mqlssltsed savyyfargr 101 pdyygsskwy fdvwgqgttv tvss; (SEQ ID NO: 67) 1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgrglewigr 51 idpnsggtky nekfkskatl tvdkpsstay mqlssltsed savyycaryd 101 yygssyfdyw gqgttvtvss; (SEQ ID NO: 68) 1 qiqlqqsgpe lvrpgasvki sckasgytft dyyihwvkqr pgeglewigw 51 iypgsgntky nekfkgkatl tvdtssstay mqlssltsed savyfcargg 101 kfamdywgqg tsvtvss; (SEQ ID NO: 69) 1 qvqlqqsgae lvkpgasvkl sckasgytft sywmhwvkqr pgqglewige 51 inpsngrtny nekfkrkatl tvdkssstay mqlssltsed savyyfargr 101 pdyygsskwy fdvwgagttv tvss; (SEQ ID NO: 70) 1 qiqlqqsgpe lvkpgasvki sckasgytft dyyinwmkqk pgqglewigw 51 idpgsgntky nekfkgkatl tvdtssstay mqlssltsed tavyfcarek 101 ttyyyamdyw gqgtsvtvsa; (SEQ ID NO: 71) 1 vqlqqsgael mkpgasvkis ckasgytfsd ywiewvkqrp ghglewigei 51 lpgsgstnyh erfkgkatft adtssstaym qlnsltseds gvyyclhgny 101 dfdgwgqgtt ltvss; and (SEQ ID NO: 72) 1 qvqllesgae lmkpgasvki sckatgytfs sfwiewvkqr pghglewige 51 ilpgsggthy nekfkgkatf tadkssntay mqlssltsed savyycargh 101 syyfydgdyw gqgtsvtvss;

[0086] and/or a light chain immunoglobulin variable region selected from the group consisting of:

TABLE-US-00028 (SEQ ID NO: 73) 1 dvlmtqipvs lpvslgdqas iscrssqiiv hnngntylew ylqkpgqspq 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp 101 ftfgsgtkle ikr; (SEQ ID NO: 74) 1 dvlmtqtpls lpvslgdpas iscrssgsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 75) 1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 76) 1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 77) 1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 78) 1 dvlmtqtpls lpvslgdqas iscrssqxiv hsngntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp 101 xtfgggtkle ikr; (SEQ ID NO: 79) 1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 80) 1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 81) 1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 82) 1 dvlmtqipvs lpvslgdqas iscrssqiiv hnngntylew ylqkpgqspq 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp 101 ftfgsgtkle ikr; (SEQ ID NO: 83) 1 dvlmtqtpls lpvslgdqas iscrfsqsiv hsngntylew ylqksgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp 101 rtfgggtkle ikr; (SEQ ID NO: 84) 1 dvlmtqtpls lpvslgdqas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 85) 1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 86) 1 elvmtqtpls lpvslgdqas iscrssqtiv hsngdtyldw flqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 87) 1 dvlmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 88) 1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 89) 1 dvlmtqtpvs lsvslgdqas iscrssqsiv hstgntylew ylqkpgqspk 51 lliykisnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqashap 101 rtfgggtkle ikr; (SEQ ID NO: 90) 1 dvlmtqtpls lpvslgdqas isckssqsiv hssgntyfew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgship 101 ftfgsgtkle ikr; (SEQ ID NO: 91) 1 dieltqtpls lpvslgdqas iscrssqsiv hsngntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp 101 ytfgggtkle ikr; (SEQ ID NO: 92) 1 dvlmtqtpls lpvslgdqas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 93) 1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspr 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 94) 1 dvlmtqtpls lpvslgdqas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 95) 1 dvvmtqtpls lpvslgdpas iscrssqsiv hsnvntylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gagtdftlri srveaedlgi yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 96) 1 dvlmtqtpls lpvslgdqas iscrsnqtil lsdgdtylew ylqkpgqspk 51 lliykvsnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgshvp 101 ptfgggtkle ikr; (SEQ ID NO: 97) 1 dvlmtqtpls lpvslgdqas iscrssqtiv hsngntylew ylqkpgqspk 51 lliykvtnrf sgvpdrfsgs gsgtdftlki srveaedlgv yycfqgthap 101 ytfgggtkle ikr; and (SEQ ID NO: 98) 1 dvlmtqtpls lpvslgdqas iscrssqsiv hsngntylew ylqkpgqspk 51 lliysissrf sgvpdrfsgs gsgtdftlki srvqaedlgv yycfqgshvp 101 ytfgggtkle ikr.

[0087] The scope of the present invention includes methods wherein a patient is administered an anti-insulin-like growth factor receptor-1 (IGF1R) antibody wherein the variable region of the antibody is linked to any immunoglobulin constant region. In an embodiment, the light chain variable region is linked to a κ chain constant region. In an embodiment, the heavy chain variable region is linked to a γ1, γ2, γ3 or γ4 chain constant region. Any of the immunoglobulin variable regions set forth herein, in embodiments of the invention, can be linked to any of the foregoing constant regions.

[0088] In an embodiment of the invention, a linear antibody is an antibody fragment as described in Zapata et al. Protein Eng. 8(10):1057-1062 (1995). Briefly, these fragments comprise a pair of tandem Fd segments (VH--CH1-VH--CH1) which form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific.

[0089] In an embodiment of the invention, single-chain Fv or sFv antibody fragments comprise the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the sFv to form the desired structure for antigen binding. For a review of sFv see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0090] In an embodiment of the invention, an immunoliposome is a liposome including an anti-IGF1R antibody or antigen-binding fragment thereof. Liposomes containing the antibody or fragment can be prepared by methods known in the art, such as described in Epstein et al., Proc. Natl. Acad. Sci. USA, 82:3688 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA, 77:4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545. Liposomes with enhanced circulation time are disclosed in U.S. Pat. No. 5,013,556. Other useful liposomes can be generated by the reverse phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter. Fab fragments of an anti-IGF1R antibody can be conjugated to the liposomes as described in Martin et al. J. Biol. Chem. 257: 286-288 (1982) via a disulfide interchange reaction.

[0091] In an embodiment of the invention, a bispecific or bifunctional antibody is an artificial hybrid antibody having two different heavy/light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods including fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai, et al., (1990) Clin. Exp. Immunol. 79: 315-321, Kostelny, et al., (1992) J. Immunol. 148:1547-1553. In addition, bispecific antibodies may be formed as "diabodies" (Holliger, et al., (1993) PNAS USA 90:6444-6448) or as "Janusins" (Traunecker, et al., (1991) EMBO J. 10:3655-3659 and Traunecker, et al., (1992) Int. J. Cancer Suppl. 7:51-52).

[0092] In an embodiment of the invention, a chimeric antibody is an antibody which comprises a variable region of the present invention fused or chimerized with an antibody region (e.g., constant region) from another, non-human species (e.g., mouse, horse, rabbit, dog, cow, chicken).

[0093] In an embodiment of the invention, a disulfide stabilized Fv fragment or dsFv is an anti-IGF1R antibody or antigen-binding fragment thereof comprising a variable heavy chain (VH) and a variable light chain (VL) which are linked by a disulfide bridge.

[0094] In an embodiment of the invention, an antigen-binding fragment is a F(ab)₂ fragments which may be produced by enzymatic cleavage of an IgG by, for example, pepsin; or a Fab fragment that may be produced by, for example, reduction of F(ab)₂ with dithiothreitol or mercaptoethylamine. In an embodiment of the invention, a Fab fragment is a VL-CL chain appended to a VH--CH1 chain by a disulfide bridge. In an embodiment of the invention, a F(ab)₂ fragment is two Fab fragments which, in turn, are appended by two disulfide bridges. The Fab portion of an F(ab)₂ molecule includes a portion of the Fc region between which disulfide bridges are located.

[0095] In an embodiment of the invention, an FV fragment is a VL or VH region of an antibody.

[0096] In an embodiment of the invention, an anti-IGFR1 antibody or antigen-binding fragment thereof is conjugated to a chemical moiety. In an embodiment of the invention, the chemical moiety is a polymer which increases the half-life of the antibody or antigen-binding fragment thereof in the body of a subject. Suitable polymers include, but are not limited to, polyethylene glycol (PEG) (e.g., PEG with a molecular weight of 2 kDa, 5 kDa, 10 kDa, 12 kDa, 20 kDa, 30 kDa or 40 kDa), dextran and monomethoxypolyethylene glycol (mPEG). Lee, et al., (1999) (Bioconj. Chem. 10:973-981) discloses PEG conjugated single-chain antibodies.

[0097] In an embodiment of the invention, an IGF1R inhibitor that is administered to a subject or patient in a method according to the invention is AEW-541 (NVP-AEW-541; NVP-AEW-541-NX-7):

##STR00014##

##STR00015##

or

[0098] In an embodiment of the invention, an IGF1R inhibitor that is administered to a patient in a method according to the invention is any IGF1R anti-sense nucleic acid. For example, in an embodiment of the invention, the anti-sense IGF1R nucleic acid is ATL-1101 (Antisense Therapeutics Ltd; Australia). In an embodiment of the invention, the IGF1R anti-sense nucleic acid comprises any of the following nucleotide sequences: 5'-ATCTCTCCGCTTCCTTTC-3' (SEQ ID NO: 99), 5'-ATCTCTCCGCTTCCTTTC-3' (SEQ ID NO: 100), 5'-ATCTCTCCGCTTCCTTTC-3' (SEQ ID NO: 101) or any IGF1R antisense nucleic acid set forth in any of US Published Patent Application No. US20030096769; Published International Application No. WO 2003/100059; Fogarty et al., Antisense Nucleic Acid Drug Dev. 2002 December; 12(6):369-77; White et al., J Invest Dermatol. 2002 June; 118(6):1003-7; White et al., Antisense Nucleic Acid Drug Dev. 2000 June; 10(3):195-203; or Wraight et al., Nat. Biotechnol. 2000 May; 18(5):521-6.

[0099] In an embodiment of the invention, an IGF1R inhibitor that is administered to a patient in a method according to the invention is an anti-IGF-I or II antibody; for example, any antibody disclosed in WO 2003/93317 or EP00492552.

[0100] The scope of the present invention includes any kinase inhibitor compound set forth in published international applications WO 2004/030627 or WO 2004/030625. In an embodiment, the kinase inhibitor is (±)-4-[2-(3-chloro-4-fluoro-phenyl)-2-hydroxy-ethylamino]-3-[6-(imidaz- ol-1-yl)-4-methyl-1H-benzimidazol-2-yl]-1H-pyridin-2-one:

##STR00016##

[0101] In an embodiment of the invention, the IGR1R inhibitor is a soluble fragment of IGF1R (e.g., amino acids 30-902 of IGF1R) or siRNA (small interfering RNA) against IGF1R.

[0102] In an embodiment, IGF1R comprises the amino acid sequence set forth under Genbank Accession No.: XM_--052648 or NM_--000612.

[0103] The present invention also includes embodiments wherein the patient receives both an IGF1R inhibitor in association with one or more other agents that are useful for the treatment or prevention of any APC-mediated medical disorder such as FAP. In an embodiment, the other agent is an anti-inflammatory agent such as a non-steroidal anti-inflammatory drug. For example, a suitable non-steroidal anti-inflammatory drug is sulindac, aptosyn (sulindac sulfone), indomethacin, rofecoxib, celecoxib or aspirin. In an embodiment of the invention, an IGF1R inhibitor is administered along with dietary calcium supplementation (e.g., 1,250 to 2,000 mg of calcium; e.g., 1500 or 3000 mg/day calcium carbonate) or dietary supplementation with retinoids, carotenoids, ascorbic acid, α-tocopherol, selenium, folate or methionine. The present invention further includes embodiment wherein two or more IGF1R inhibitors are administered in association with one another.

[0104] The present invention also comprise embodiments wherein a subject is administered an IGF1R inhibitor to treat or prevent any APC-mediated medical disorder such as FAP in association with a medical therapeutic or diagnostic procedure. For example, an IGF1R inhibitor can be administered in association with colectomy (e.g., total colectomy with mucosal protectomy with ileo-anal pull through or subtotal colectomy with ileorectal anastomosis), surgical osteoma removal, endoscopic or surgical removal of adenomas (e.g., duodenal adenomas), screening for hepatoblastoma by ultrasound examination, measurement of serum alpha-fetoprotein concentration, sigmoidoscopy, colonoscopy, esophagogastroduodenoscopy or small bowel X-ray.

[0105] The term "in association" indicates that the components of the combinations of the invention can be formulated into a single composition for simultaneous delivery or formulated separately into two or more compositions (e.g., a kit). Moreover, compositions and procedures making up combinations of the invention can be administered simultaneously or non-simultaneously. Furthermore, each component of a combination of the invention can be administered to a subject at a different time than when the other component is administered and in any order or permutation; for example, each administration may be given non-simultaneously at several intervals over a given period of time. Moreover, the separate components may be administered to a subject by the same or by a different route (e.g., orally, intravenously, intratumorally).

[0106] The scope of the present invention encompasses compositions comprising any IGF1R inhibitor (e.g., as set forth herein) in association with any chemotherapeutic agent (e.g., as set forth herein) known to be useful for treating or preventing any APC-mediated medical disorder such as FAP (e.g., celecoxib) along with pharmaceutical compositions thereof.

Generation of Antibodies

[0107] Any suitable method can be used to elicit an antibody or antigen-binding fragment thereof with the desired biologic properties to inhibit IGF1R. It is desirable to prepare monoclonal antibodies (mAbs) from various mammalian hosts, such as mice, rodents, primates, humans, etc. Description of techniques for preparing such monoclonal antibodies may be found in, e.g., Stites, et al. (eds.) BASIC AND CLINICAL IMMUNOLOGY (4th ed.) Lange Medical Publications, Los Altos, Calif., and references cited therein; Harlow and Lane (1988) ANTIBODIES: A LABORATORY MANUAL CSH Press; Goding (1986) MONOCLONAL ANTIBODIES: PRINCIPLES AND PRACTICE (2d ed.) Academic Press, New York, N.Y. Thus, monoclonal antibodies may be obtained by a variety of techniques familiar to researchers skilled in the art. Typically, spleen cells from an animal immunized with a desired antigen are immortalized, commonly by fusion with a myeloma cell. See Kohler and Milstein (1976) Eur. J. Immunol. 6:511-519. Alternative methods of immortalization include transformation with Epstein Barr Virus, oncogenes, or retroviruses, or other methods known in the art. See, e.g., Doyle, et al. (eds. 1994 and periodic supplements) CELL AND TISSUE CULTURE: LABORATORY PROCEDURES, John Wiley and Sons, New York, N.Y. Colonies arising from single immortalized cells are screened for production of antibodies of the desired specificity and affinity for the antigen, and yield of the monoclonal antibodies produced by such cells may be enhanced by various techniques, including injection into the peritoneal cavity of a vertebrate host. Alternatively, one may isolate DNA sequences which encode a monoclonal antibody or an antigen-binding fragment thereof by screening a DNA library from human B cells according, e.g., to the general protocol outlined by Huse, et al. (1989) Science 246:1275-1281. Modified antibodies can be generated, for example, by introducing mutations in DNA encoding an immunoglobulin chain, for example, by use of conventional recombinant biological techniques.

[0108] Other suitable techniques involve selection of libraries of antibodies in phage or similar vectors. See, e.g., Huse et al., Science 246:1275-1281 (1989); and Ward et al., Nature 341:544-546 (1989).

[0109] In an embodiment of the invention, recombinant immunoglobulins are produced, see Cabilly U.S. Pat. No. 4,816,567; and Queen et al. (1989) Proc. Nat'l Acad. Sci. USA 86:10029-10033; or made in transgenic mice, see Mendez et al. (1997) Nature Genetics 15:146-156. Further methods for producing chimeric, humanized and human antibodies are well known in the art. See, e.g., U.S. Pat. No. 5,530,101, issued to Queen et al, U.S. Pat. No. 5,225,539, issued to Winter et al, U.S. Pat. Nos. 4,816,397 issued to Boss et al., all of which are incorporated by reference in their entirety.

[0110] In an embodiment of the invention, a fully-human monoclonal antibody directed against IGF1R is generated using transgenic mice carrying parts of the human immune system rather than the mouse system. These transgenic mice, which may be referred to, herein, as "HuMAb" mice, contain a human immunoglobulin gene miniloci that encodes unrearranged human heavy (μ and γ) and κ light chain immunoglobulin sequences, together with targeted mutations that inactivate the endogenous μ and κ chain loci (see e.g., Lonberg N et al., (1994) Nature 368(6474): 856-859; Lonberg N, et al., (1994), supra; reviewed in Lonberg, N. (1994) Handbook of Experimental Pharmacology 113:49-101; Lonberg, N., et al., (1995) Intern. Rev. Immunol. 13:65-93, and Harding, F., et al., (1995) Ann. N.Y. Acad. Sci. 764:536-546; in Taylor, L., et al., (1992) Nucleic Acids Research 20:6287-6295; Chen, J., et al., (1993) International Immunology 5: 647-656; Tuaillon, et al., (1993) Proc. Natl. Acad. Sci. USA 90:3720-3724; Choi, et al., (1993) Nature Genetics 4:117-123; Chen, J., et al., (1993) EMBO J. 12: 821-830; Tuaillon, et al., (1994) J. Immunol. 152:2912-2920; Lonberg, et al., (1994) Nature 368(6474): 856-859; Lonberg, N. (1994) Handbook of Experimental Pharmacology 113:49-101; Taylor, L., et al., (1994) International Immunology 6: 579-591; Lonberg, N., et al., (1995) Intern. Rev. Immunol. Vol. 13: 65-93; Harding, F., et al., (1995) Ann. N.Y. Acad. Sci. 764:536-546; Fishwild, D., et al., (1996) Nature Biotechnology 14: 845-851 and Harding, et al., (1995) Annals NY Acad. Sci. 764:536-546; U.S. Pat. Nos. 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,789,650; 5,877,397; 5,661,016; 5,814,318; 5,874,299; 5,770,429 and 5,545,807; and International Patent Application Publication Nos. WO 98/24884; WO 94/25585; WO 93/12227; WO 92/22645 and WO 92/03918).

[0111] A plasmid system set forth in published U.S. patent application no. US2005/0176099 is also useful of the generation of an anti-IGF1R antibody of the present invention (e.g., LCF/HCA).

Dosage

[0112] In an embodiment of the invention, an IGF1R inhibitor is administered to a patient at a "therapeutically effective dosage" or "therapeutically effective amount" which inhibits an APC-mediated disease or condition (e.g., FAP) to any extent--e.g., by at least about 20%, 40%, 60% or 80%-100% relative to untreated subjects.

[0113] In an embodiment of the invention, the term "therapeutically effective amount" or "therapeutically effective dosage" means that amount or dosage of an IGF1R inhibitor (e.g., an anti-IGF1R antibody or antigen-binding fragment thereof) that will elicit a biological or medical response of a tissue, system, subject or host that is being sought by the administrator (such as a researcher, doctor or veterinarian) which includes any measurable alleviation of the signs, symptoms and/or clinical indicia of an APC-mediated disease (e.g., FAP) and/or the prevention, slowing or halting of progression of FAP to any degree.

[0114] A physician or veterinarian having ordinary skill in the art may determine and prescribe the effective amount of IGF1R inhibitor required for an optimal clinical result depending on the circumstances of the particular patient. For example, the physician or veterinarian could start doses of an IGF1R inhibitor employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. The effectiveness of a given dose or treatment regimen of IGF1R inhibitor can be determined, for example, by consideration of any of several observed clinical indicia. Proper adjustments to dosage and/or regimen can then be made by the clinician so as to reach the best result. For example, a clinician can monitor the responsiveness of an APC-mediated disease (e.g., FAP, Gardner Syndrome or Turcot Syndrome) to a given course of treatment by any appropriate means. For example, responsiveness can be monitored by performing sigmoidoscopy or colonoscopy to visualize the condition of the subject's colon. A clinician can monitor the state of any upper gastrointestinal tract polyps (e.g., in the stomach, duodenum or papilla), in a subject, by way of an upper endoscopic examination (e.g., a forward and a side viewing endoscopic examination). Another monitoring method includes patient interviews to determine the presence and severity of any symptoms associated with FAP, for example, blood in the stool, diarrhea that is not the result of diet or illness, constipation, crampy pain in the abdomen, irregular bowel habits, decrease in the size or caliber of stool, frequent feeling of distention in the abdomen or bowel region (e.g., gas pain, bloating, fullness, with or without cramping), unexplained weight loss, vomiting and lack of energy.

[0115] The presence of any of the foregoing symptoms and clinical indicia can also aid a practitioner in identifying a patient with an APC-mediated medical disorder, such as FAP, who is thus in need of a treatment of the present invention.

[0116] As stated above, the present invention comprises methods for treatment or prevention of any medical condition mediated by a mutation in the APC gene, for example, FAP, by administration of a "therapeutically effective amount" or "therapeutically effective dose" of an IGF1R inhibitor. In an embodiment of the invention, a therapeutically effective daily amount or dose of IGF1R inhibitor or pharmaceutical composition thereof is administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day. In an embodiment, a "therapeutically effective" dosage or amount of any anti-IGF1R antibody (e.g., 19D12/15H12 LCD/HCB, LCC/HCB or LCF/HCA) is in the range of about 0.3 mg/kg (body weight) to about 20 mg/kg (e.g., 0.5 mg/kg, 0.9 mg/kg, 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg/kg, 8 mg/kg, 9 mg/kg, 10 mg/kg, 11 mg/kg, 12 mg/kg, 13 mg/kg, 14 mg/kg, 15 mg/kg, 16 mg/kg, 17 mg/kg, 18 mg/kg, 19 mg/kg or 20 mg/kg) administered, for example, 1 time per week. In an embodiment of the invention, a therapeutically effective dosage or amount of an IGF1R inhibitor or any other chemotherapeutic agent (e.g., as set forth herein) is, whenever possible, as set forth in the Physicians' Desk Reference 2003 (Thomson Healthcare; 57^th edition (Nov. 1, 2002)) which is herein incorporated by reference or in the scientific literature. For example, in an embodiment of the invention, a therapeutically effective dosage or amount of celecoxib is about 400 mg twice per day taken with food. In an embodiment of the invention, a therapeutically effective dosage or amount of sulindac is about 400 mg per day. In an embodiment of the invention, a therapeutically effective dosage or amount of sulindac sulfone is about 200 mg/day.

Therapeutic Methods and Administration

[0117] The term "patient" or "subject" includes any organism, preferably an animal, for example, a mammal (e.g., rat, mouse, dog, cat, rabbit) such as a human. Accordingly, the present invention includes veterinary and non-veterinary methods for treating or preventing an APC-mediated disease.

[0118] As stated above, in an embodiment of the invention, where possible, an IGF1R inhibitor or any other chemotherapeutic agent set forth herein (e.g., NSAID) is administered to a patient or subject in accordance with the Physicians' Desk Reference 2003 (Thomson Healthcare; 57th edition (Nov. 1, 2002)) or as set forth herein.

[0119] An IGF1R inhibitor and/or an additional therapeutic agent (e.g., an NSAID such as celecoxib) can be administered by an invasive route such as by injection. In an embodiment of the invention, an anti-IGF1R antibody, or pharmaceutical composition thereof, is administered intravenously, subcutaneously, intramuscularly or intraarterially. An embodiment of the invention comprises treating or preventing an APC-mediated medical condition by reconstituting a lyophilized or otherwise desiccated powder containing an IGF1R inhibitor (e.g., a pharmaceutical formulation thereof, e.g., in a glass or plastic vial or container), for example in water, DMSO or buffer, and then administering a therapeutically effective amount of the agent in the reconstituted solution to a subject in need thereof (e.g., a subject suffering from FAP), for example, parenterally (e.g., by injection with a hypodermic needle).

[0120] Administration of an IGF1R inhibitor and/or an additional therapeutic agent by a non-invasive route (e.g., orally; for example, in a pill, capsule or tablet) is also within the scope of the present invention.

[0121] The scope of the present invention comprises methods wherein the IGF1R inhibitor is administered by the same or by a different route as an additional therapeutic agent (e.g., celecoxib). In an embodiment of the invention, an anti-IGF1R antibody is administered by injection and a second therapeutic agent (e.g., celecoxib) is administered orally.

[0122] An IGF1R inhibitor and/or an additional therapeutic agent can be administered with medical devices known in the art. For example, a pharmaceutical composition of the invention (e.g., a pharmaceutical composition comprising an anti-IGF1R antibody) can be administered by injection with a hypodermic needle.

[0123] The pharmaceutical compositions of the invention may also be administered with a needleless hypodermic injection device; such as the devices disclosed in U.S. Pat. Nos. 6,620,135; 6,096,002; 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824 or 4,596,556.

[0124] Examples of well-known implants and modules for administering pharmaceutical compositions include: U.S. Pat. No. 4,487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Pat. No. 4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; U.S. Pat. No. 4,447,224, which discloses a variable flow implantable infusion apparatus for continuous drug delivery; U.S. Pat. No. 4,439,196, which discloses an osmotic drug delivery system having multi-chamber compartments. Many other such implants, delivery systems, and modules are well known to those skilled in the art. Diagnosis of a medical disorder such as FAP, Gardner Syndrome, Turcot Syndrome and Attenuated FAP is within the scope of knowledge commonly held in the art. For example, Gardner Syndrome can be diagnosed using radiological tests which reveal long bone osteomas and subtle defects in the mandible; eye exams that reveal pigmented lesions of the fundus of the eye; and colonoscopies and other invasive tests which reveal the presence of polyps. FAP and AFAP can be diagnosed by screening of the patient's colon for the presence of polyps (e.g., by flexible sigmoidoscopy or colonoscopy) as well as by genetic screening for the presence of a mutated APC gene.

Pharmaceutical Compositions

[0125] An IGF1R inhibitor can, in an embodiment of the invention, be incorporated into a pharmaceutical composition, along with a pharmaceutically acceptable carrier, suitable for administration to a subject in vivo is within the scope of the present invention. The scope of the present invention includes pharmaceutical compositions which are suitable to be administered to a subject by any route (e.g., parenteral or non-parenteral) including, for example, oral, ocular, topical, pulmonary (inhalation), intratumoral injection, intravenous injection, subcutaneous injection or intramuscular injection.

[0126] For general information concerning formulations, see, e.g., Gilman, et al., (eds.) (1990), The Pharmacological Bases of Therapeutics, 8th Ed., Pergamon Press; A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co., Easton, Pa.; Avis, et al., (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications Dekker, New York; Lieberman, et al., (eds.) (1990) Pharmaceutical Dosage Forms: Tablets Dekker, New York; and Lieberman, et al., (eds.) (1990), Pharmaceutical Dosage Forms: Disperse Systems Dekker, New York, Kenneth A. Walters (ed.) (2002) Dermatological and Transdermal Formulations (Drugs and the Pharmaceutical Sciences), Vol 119, Marcel Dekker.

[0127] Pharmaceutically acceptable carriers are conventional and very well known in the art. Examples include aqueous and nonaqueous carriers, stabilizers, antioxidants, solvents, dispersion media, coatings, antimicrobial agents, buffers, serum proteins, isotonic and absorption delaying agents, and the like that are physiologically compatible. In an embodiment of the invention, the carrier is suitable for injection into a subject's body.

[0128] Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0129] Examples of pharmaceutically-acceptable antioxidants include: water soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; and oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0130] Prevention of the presence of microorganisms may be ensured both by sterilization procedures, and by the inclusion of various antimicrobial agents such as EDTA, EGTA, paraben, chlorobutanol, phenol sorbic acid, and the like.

[0131] Suitable buffers which may be included in the pharmaceutical compositions of the invention include L-histidine-based buffers, phosphate-based buffers (e.g., phosphate buffered saline, pH≈7), sorbate-based buffers or glycine-based buffers.

[0132] Serum proteins which are, in an embodiment of the invention, included in the pharmaceutical compositions of the invention include human serum albumin.

[0133] Isotonic agents, such as sugars (e.g., sucrose), ethanol, polyalcohols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, mannitol or sorbitol), sodium citrate or sodium chloride (e.g., buffered saline) are, in an embodiment of the invention, included in the pharmaceutical compositions of the invention. In an embodiment of the invention, the sugar, for example, glucose or sucrose is present at a high concentration (e.g., about 10-100 mg/ml, e.g., 50 mg/ml, 60 mg/ml or 70 mg/ml).

[0134] Prolonged absorption of an injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and/or gelatin.

[0135] Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils.

[0136] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is well known in the art.

[0137] Sterile injectable solutions comprising an anti-IGF1R antibody can be prepared by incorporating the antibody or antigen-binding fragment thereof in the required amount in an appropriate solvent, optionally with one or a combination of ingredients enumerated above, as required, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporating the antibody into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient plus any additional, desired ingredient from a previously sterile-filtered solution thereof.

[0138] In an embodiment of the invention, an anti-IGF1R antibody or antigen-binding fragment thereof of the invention is in a pharmaceutical formulation comprising a therapeutically effective amount of said antibody or fragment, a buffer and sucrose. For example, in an embodiment of the invention, the buffer is any one of phosphate buffer, citrate buffer, histidine buffer, glycine buffer or acetate buffer. The pharmaceutical formulation can be within any suitable pH range. In an embodiment of the invention, the pH is 5.0, 5.5, 6.0, 7.5, or between about 5.5 and about 6 or between about 5 and about 7.

[0139] In an embodiment of the invention, an IGF1R inhibitor and/or other chemotherapeutic agent is orally administered. Pharmaceutical compositions for oral administration contain, in an embodiment of the invention, in addition to the active agent, additives such as starch (e.g., potato, maize or wheat starch or cellulose), starch derivatives (e.g., microcrystalline cellulose or silica), sugars (e.g., lactose), talc, stearate, magnesium carbonate or calcium phosphate. In order to ensure that oral compositions comprising an active agent are well tolerated by the patient's digestive system, mucus formers or resins may be included. It may also be desirable to improve tolerance by formulating the active agent in a capsule which is insoluble in the gastric juices. An exemplary pharmaceutical composition of this invention in the form of a capsule is prepared by filling a standard two-piece hard gelatin capsule with the active agent in powdered form, lactose, talc and magnesium stearate.

[0140] Oral administration of immunoglobulins, such as an anti-IGF1R antibody or antigen-binding fragment thereof of the invention, has been described (Foster, et al., (2001) Cochrane Database System rev. 3:CD001816).

[0141] In an embodiment of the invention, an anti-IGF1R antibody or antigen-binding fragment thereof of the invention is in a pharmaceutical formulation suitable for injection comprising, at a pH of about 5.5, about 20 mg/ml of the antibody (or any suitable concentration of the antibody); about 2.3 mg/ml of sodium acetate trihydrate; about 0.18 mg/ml of glacial acetic acid; about 70 mg/ml of sucrose and water.

[0142] An IGF1R inhibitor or other chemotherapeutic agent may also be administered by inhalation. A suitable pharmaceutical composition for inhalation may be an aerosol. An exemplary pharmaceutical composition for inhalation of an IGF1R inhibitor or other chemotherapeutic agent may include: an aerosol container with a capacity of 15-20 ml comprising the IGF1R inhibitor or other chemotherapeutic agent, a lubricating agent, such as polysorbate 85 or oleic acid, dispersed in a propellant, such as freon, preferably in a combination of 1,2-dichlorotetrafluoroethane and difluorochloromethane. Preferably, the composition is in an appropriate aerosol container adapted for either intranasal or oral inhalation administration.

EXAMPLES

[0143] The following information is provided for more clearly describing the present invention and should not be construed to limit the present invention. Any and all of the compositions and methods described below fall within the scope of the present invention.

Example 1

Inhibition of Polyposis in Human IGF1R Knock-in Mice

[0144] In this example, the ability of anti-IGF1R antibody 19D12 (LCF/HCA: amino acids 20-128 of SEQ ID NO: 8 and 20-137 of SEQ ID NO: 10) to treat familial adenomatous polyposis (FAP) is established in a mouse model for the disease. The model comprises the Apc^Min/+ allele which imparts the FAP phenotype. A phenotype that results from the allele is increased development of intestinal adenomas. The Min mutant mouse has an autosomal dominant heterozygous nonsense mutation in the mouse Apc gene (see e.g., Su et al., Science (Wash D.C.) 256: 668-670 (1992)).

[0145] The Min mice are also engineered, by a knock-in mutation, to express the human IGF1R gene. Expression of the IGF1R gene, in the Min mouse, allows determination of the ability of an anti-IGF1R antibody to alleviate the FAP phenotype. Antibody-dependent reduction of the incidence of intestinal adenomas in the hIGF1R, Apc^Min/+ mice indicates that the antibody would be an effective treatment of FAP in humans and other animals.

[0146] Twenty eight C57BI/6J (hIGF1R, Apc^Min/+) mice are obtained commercially and broken into the following groups (4 per group):

TABLE-US-00029 TABLE 1 Organization of Treatment Groups. Treatment group Route and dosing schedule NT vehicle injection once a week 19D12 (LCF/HCA), 0.02 mg/mouse i.p. injection once a week 19D12 (LCF/HCA), 0.1 mg/mouse i.p. injection once a week 19D12 (LCF/HCA), 0.5 mg/mouse i.p. injection once a week Celecoxib, 150 ppm in diet Celecoxib, 500 ppm in diet Celecoxib, 1500 ppm in diet

[0147] Small intestinal adenomas and colonic adenomas are scored for number and diameter at postnatal day 80. The stomach, small intestine, and colon are dissected free of mesentery and opened along the longitudinal axis. Intestinal contents are cleared with phosphate buffered saline (PBS), and the small intestine is divided into three equal-length segments and laid open with the colon on the absorptive side of Benchkote (Whatman). Intestines are fixed in 4% (4 g/100 ml) paraformaldehyde in PBS (24 h) followed by 70% ethanol (v/v). Using a dissecting microscope (×10-30) and calipers, adenoma number and diameter are obtained for the entire length of the small intestine and colon. Adenoma analysis is performed without knowledge of genotype by one person and confirmed independently by another. Small intestine surface area is calculated by summation of the multiples of length of each fixed segment by width, which is measured at the midpoint of each segment. Colon surface area is calculated by multiplying the length of the fixed material from the anorectal junction to the point of insertion of the small intestine, omitting the appendix, by the width at the midpoint. Raw numbers of adenomas observed as well as adenomas per unit of small intestine and colon surface area are calculated.

[0148] Mice receiving the 19D12 (LCF/HCA) antibody or celecoxib exhibit a dose-dependent reduction of raw adenoma number as well as adenomas per unit surface area in the small intestine and colon.

[0149] The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and the accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0150] Patents, patent applications, Genbank Accession Numbers and publications are cited throughout this application, the disclosures of which are incorporated herein by reference in their entireties.

Sequence CWU 1

1

1121384DNAArtificial SequenceModified 19D12/15H12 Light Chain-C 1atg tcg cca tca caa ctc att ggg ttt ctg ctg ctc tgg gtt cca gcc 48Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 tcc agg ggt gaa att gtg ctg act cag agc cca gac tct ctg tct gtg 96Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ser Val 20 25 30 act cca ggc gag aga gtc acc atc acc tgc cgg gcc agt cag agc att 144Thr Pro Gly Glu Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile 35 40 45 ggt agt agc tta cac tgg tac cag cag aaa cca ggt cag tct cca aag 192Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys 50 55 60 ctt ctc atc aag tat gca tcc cag tcc ctc tca ggg gtc ccc tcg agg 240Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Val Pro Ser Arg 65 70 75 80 ttc agt ggc agt gga tct ggg aca gat ttc acc ctc acc atc agt agc 288Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 85 90 95 ctc gag gct gaa gat gct gca gcg tat tac tgt cat cag agt agt cgt 336Leu Glu Ala Glu Asp Ala Ala Ala Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 tta cct cac act ttc ggc caa ggg acc aag gtg gag atc aaa cgt acg 384Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 2128PRTArtificial SequenceSynthetic Construct 2Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ser Val 20 25 30 Thr Pro Gly Glu Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile 35 40 45 Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys 50 55 60 Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Val Pro Ser Arg 65 70 75 80 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 85 90 95 Leu Glu Ala Glu Asp Ala Ala Ala Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 3384DNAArtificial SequenceModified 19D12/15H12 Light Chain-D 3atg tcg cca tca caa ctc att ggg ttt ctg ctg ctc tgg gtt cca gcc 48Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 tcc agg ggt gaa att gtg ctg act cag agc cca gac tct ctg tct gtg 96Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ser Val 20 25 30 act cca ggc gag aga gtc acc atc acc tgc cgg gcc agt cag agc att 144Thr Pro Gly Glu Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile 35 40 45 ggt agt agc tta cac tgg tac cag cag aaa cca ggt cag tct cca aag 192Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys 50 55 60 ctt ctc atc aag tat gca tcc cag tcc ctc tca ggg gtc ccc tcg agg 240Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Val Pro Ser Arg 65 70 75 80 ttc agt ggc agt gga tct ggg aca gat ttc acc ctc acc atc agt agc 288Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 85 90 95 ctc gag gct gaa gat ttc gca gtg tat tac tgt cat cag agt agt cgt 336Leu Glu Ala Glu Asp Phe Ala Val Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 tta cct cac act ttc ggc caa ggg acc aag gtg gag atc aaa cgt acg 384Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 4128PRTArtificial SequenceSynthetic Construct 4Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ser Val 20 25 30 Thr Pro Gly Glu Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile 35 40 45 Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys 50 55 60 Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Val Pro Ser Arg 65 70 75 80 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser 85 90 95 Leu Glu Ala Glu Asp Phe Ala Val Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 5384DNAArtificial SequenceModified 19D12/15H12 Light Chain-E 5atg tcg cca tca caa ctc att ggg ttt ctg ctg ctc tgg gtt cca gcc 48Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 tcc agg ggt gaa att gtg ctg act cag agc cca ggt acc ctg tct gtg 96Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Val 20 25 30 tct cca ggc gag aga gcc acc ctc tcc tgc cgg gcc agt cag agc att 144Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 35 40 45 ggt agt agc tta cac tgg tac cag cag aaa cca ggt cag gct cca agg 192Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 50 55 60 ctt ctc atc aag tat gca tcc cag tcc ctc tca ggg atc ccc gat agg 240Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Ile Pro Asp Arg 65 70 75 80 ttc agt ggc agt gga tct ggg aca gat ttc acc ctc acc atc agt aga 288Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 85 90 95 ctg gag cct gaa gat gct gca gcg tat tac tgt cat cag agt agt cgt 336Leu Glu Pro Glu Asp Ala Ala Ala Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 tta cct cac act ttc ggc caa ggg acc aag gtg gag atc aaa cgt aca 384Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 6128PRTArtificial SequenceSynthetic Construct 6Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Val 20 25 30 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 35 40 45 Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 50 55 60 Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Ile Pro Asp Arg 65 70 75 80 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 85 90 95 Leu Glu Pro Glu Asp Ala Ala Ala Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 7384DNAArtificial Sequence19D12/15H12 Light Chain-F 7atg tcg cca tca caa ctc att ggg ttt ctg ctg ctc tgg gtt cca gcc 48Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 tcc agg ggt gaa att gtg ctg act cag agc cca ggt acc ctg tct gtg 96Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Val 20 25 30 tct cca ggc gag aga gcc acc ctc tcc tgc cgg gcc agt cag agc att 144Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 35 40 45 ggt agt agc tta cac tgg tac cag cag aaa cca ggt cag gct cca agg 192Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 50 55 60 ctt ctc atc aag tat gca tcc cag tcc ctc tca ggg atc ccc gat agg 240Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Ile Pro Asp Arg 65 70 75 80 ttc agt ggc agt gga tct ggg aca gat ttc acc ctc acc atc agt aga 288Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 85 90 95 ctg gag cct gaa gat ttc gca gtg tat tac tgt cat cag agt agt cgt 336Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 tta cct cac act ttc ggc caa ggg acc aag gtg gag atc aaa cgt aca 384Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 8128PRTArtificial SequenceSynthetic Construct 8Met Ser Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala 1 5 10 15 Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Val 20 25 30 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile 35 40 45 Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg 50 55 60 Leu Leu Ile Lys Tyr Ala Ser Gln Ser Leu Ser Gly Ile Pro Asp Arg 65 70 75 80 Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg 85 90 95 Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys His Gln Ser Ser Arg 100 105 110 Leu Pro His Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr 115 120 125 9411DNAArtificial Sequence19D12/15H12 heavy chain-A 9atg gag ttt ggg ctg agc tgg gtt ttc ctt gtt gct ata tta aaa ggt 48Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Ile Leu Lys Gly 1 5 10 15 gtc cag tgt gag gtt cag ctg gtg cag tct ggg gga ggc ttg gta aag 96Val Gln Cys Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys 20 25 30 cct ggg ggg tcc ctg aga ctc tcc tgt gca gcc tct gga ttc acc ttc 144Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 agt agc ttt gct atg cac tgg gtt cgc cag gct cca gga aaa ggt ctg 192Ser Ser Phe Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 gag tgg ata tca gtt att gat act cgt ggt gcc aca tac tat gca gac 240Glu Trp Ile Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp 65 70 75 80 tcc gtg aag ggc cga ttc acc atc tcc aga gac aat gcc aag aac tcc 288Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser 85 90 95 ttg tat ctt caa atg aac agc ctg aga gcc gag gac act gct gtg tat 336Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 100 105 110 tac tgt gca aga ctg ggg aac ttc tac tac ggt atg gac gtc tgg ggc 384Tyr Cys Ala Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly 115 120 125 caa ggg acc acg gtc acc gtc tcc tca 411Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 10137PRTArtificial SequenceSynthetic Construct 10Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Ser Ser Phe Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Ile Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp 65 70 75 80 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser 85 90 95 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 100 105 110 Tyr Cys Ala Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly 115 120 125 Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 11411DNAArtificial SequenceModified 19D12/15H12 heavy chain-B 11atg gag ttt ggg ctg agc tgg gtt ttc ctt gtt gct ata tta aaa ggt 48Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Ile Leu Lys Gly 1 5 10 15 gtc cag tgt gag gtt cag ctg gtg cag tct ggg gga ggc ttg gta cag 96Val Gln Cys Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln 20 25 30 ccc ggg ggg tcc ctg aga ctc tcc tgt gca gcc tct gga ttc acc ttc 144Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 agt agc ttt gct atg cac tgg gtt cgc cag gct cca gga aaa ggt ctg 192Ser Ser Phe Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 gag tgg ata tca gtt att gat act cgt ggt gcc aca tac tat gca gac 240Glu Trp Ile Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp 65 70 75 80 tcc gtg aag ggc cga ttc acc atc tcc aga gac aat gcc aag aac tcc 288Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser 85 90 95 ttg tat ctt caa atg aac agc ctg aga gcc gag gac act gct gtg tat 336Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 100 105 110 tac tgt gca aga ctg ggg aac ttc tac tac ggt atg gac gtc tgg ggc 384Tyr Cys Ala Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly 115 120 125 caa ggg acc acg gtc acc gtc tcc tca 411Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 12137PRTArtificial SequenceSynthetic Construct 12Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Ser Ser Phe Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Ile Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp 65 70 75 80 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser 85 90 95 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 100 105 110 Tyr Cys Ala Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly 115 120 125 Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 13174PRTArtificial Sequenceimmunoglobulin heavy chain variable region 13Gly Arg Leu Gly Gln Ala Trp Arg Ser Leu Arg Leu Ser Cys Ala Ala 1 5 10 15 Ser Gly Phe Thr Phe Ser Asp Tyr Tyr Met Ser Trp Ile Arg Gln Ala 20 25 30 Pro Gly Lys Gly Leu Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser 35 40 45 Thr Arg Asp Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile

Ser Arg 50 55 60 Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala 65 70 75 80 Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg Asp Gly Val Glu Thr Thr 85 90 95 Phe Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ser Cys Ala 165 170 14124PRTArtificial Sequenceimmunoglobulin heavy chain variable region 14Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser 1 5 10 15 Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ala 20 25 30 Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 35 40 45 Ala Ile Ser Gly Ser Gly Gly Thr Thr Phe Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Thr Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Asp Leu Gly Trp Ser Asp Ser Tyr Tyr Tyr Tyr Tyr Gly Met Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 15112PRTArtificial Sequenceimmunoglobulin heavy chain variable region 15Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr 1 5 10 15 Val Ser Gly Gly Ser Ile Ser Asn Tyr Tyr Trp Ser Trp Ile Arg Gln 20 25 30 Pro Ala Gly Lys Gly Leu Glu Trp Ile Gly Arg Ile Tyr Thr Ser Gly 35 40 45 Ser Pro Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Met Ser Val 50 55 60 Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Asn Ser Val Thr Ala 65 70 75 80 Ala Asp Thr Ala Val Tyr Tyr Cys Ala Val Thr Ile Phe Gly Val Val 85 90 95 Ile Ile Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 100 105 110 16125PRTArtificial Sequenceimmunoglobulin heavy chain variable region 16Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ile Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Asp Leu Gly Tyr Gly Asp Phe Tyr Tyr Tyr Tyr Tyr Gly Met 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 17113PRTArtificial Sequenceimmunoglobulin heavy chain variable region 17Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val 1 5 10 15 Ser Gly Gly Ser Ile Ser Ser Tyr Tyr Trp Ser Trp Ile Arg Gln Pro 20 25 30 Pro Gly Lys Gly Leu Glu Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser 35 40 45 Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Asp 50 55 60 Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala 65 70 75 80 Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Tyr Ser Ser Ser Phe Tyr 85 90 95 Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 100 105 110 Ser 18122PRTArtificial Sequenceimmunoglobulin heavy chain variable region 18Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Thr Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Asp Pro Gly Thr Thr Val Ile Met Ser Trp Phe Asp Pro Trp 100 105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 19136PRTArtificial Sequenceimmunoglobulin light chain variable region 19Ala Ser Val Gly Asp Arg Val Thr Phe Thr Cys Arg Ala Ser Gln Asp 1 5 10 15 Ile Arg Arg Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 20 25 30 Lys Arg Leu Ile Tyr Ala Ala Ser Arg Leu Gln Ser Gly Val Pro Ser 35 40 45 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 50 55 60 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn 65 70 75 80 Asn Tyr Pro Arg Thr Phe Gly Gln Gly Thr Glu Val Glu Ile Ile Arg 85 90 95 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 100 105 110 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 115 120 125 Pro Arg Glu Ala Lys Val Gln Trp 130 135 20107PRTArtificial Sequenceimmunoglobulin light chain variable region 20Asp Ile Gln Met Thr Gln Phe Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Arg Leu His Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Cys 85 90 95 Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 21100PRTArtificial Sequenceimmunoglobulin light chain variable region 21Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Phe Thr Cys Arg 1 5 10 15 Ala Ser Gln Asp Ile Arg Arg Asp Leu Gly Trp Tyr Gln Gln Lys Pro 20 25 30 Gly Lys Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Arg Leu Gln Ser 35 40 45 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr 50 55 60 Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 65 70 75 80 Leu Gln His Asn Asn Tyr Pro Arg Thr Phe Gly Gln Gly Thr Glu Val 85 90 95 Glu Ile Ile Arg 100 22107PRTArtificial Sequenceimmunoglobulin light chain variable region 22Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Ser Asp 20 25 30 Leu Gly Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Lys Leu His Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Arg Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 2392PRTArtificial Sequenceimmunoglobulin light chain variable region 23Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Thr 1 5 10 15 Phe Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu 20 25 30 Ile His Val Ala Ser Ser Leu Gln Gly Gly Val Pro Ser Arg Phe Ser 35 40 45 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln 50 55 60 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Asn Ala Pro 65 70 75 80 Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 85 90 2491PRTArtificial Sequenceimmunoglobulin light chain variable region 24Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Gly Arg Tyr 1 5 10 15 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 20 25 30 Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 35 40 45 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro 50 55 60 Glu Asp Phe Ala Val Phe Tyr Cys Gln Gln Tyr Gly Ser Ser Pro Arg 65 70 75 80 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 85 90 25236PRTArtificial Sequencelight chain immunoglobulin 25Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Gly Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110 His Asn Ser Tyr Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 26236PRTArtificial Sequencelight chain immunoglobulin 26Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Phe Thr Cys Arg Ala Ser 35 40 45 Gln Asp Ile Arg Arg Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Arg Leu Gln Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110 His Asn Asn Tyr Pro Arg Thr Phe Gly Gln Gly Thr Glu Val Glu Ile 115 120 125 Ile Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 27236PRTArtificial Sequencelight chain immunoglobulin 27Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Gly Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110 His Asn Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 28236PRTArtificial Sequencelight chain immunoglobulin 28Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Phe Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Gly Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Arg Leu His Arg Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100

105 110 His Asn Ser Tyr Pro Cys Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 29473PRTArtificial Sequenceheavy chain immunoglobulin 29Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Ile Ile Lys Gly 1 5 10 15 Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Ser Asp Tyr Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 85 90 95 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Val Leu Arg Phe Leu Glu Trp Leu Leu Tyr Tyr 115 120 125 Tyr Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr 130 135 140 Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 145 150 155 160 Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val 165 170 175 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 180 185 190 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 195 200 205 Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly 210 215 220 Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys 225 230 235 240 Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys 245 250 255 Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 260 265 270 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 275 280 285 Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 290 295 300 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 305 310 315 320 Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His 325 330 335 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 340 345 350 Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln 355 360 365 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met 370 375 380 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 385 390 395 400 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 405 410 415 Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu 420 425 430 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 435 440 445 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 450 455 460 Lys Ser Leu Ser Leu Ser Pro Gly Lys 465 470 30470PRTArtificial Sequenceheavy chain immunoglobulin 30Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Ile Ile Lys Gly 1 5 10 15 Val Gln Cys Gln Ala Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Ser Asp Tyr Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser Thr Arg Asp Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 85 90 95 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Val Arg Asp Gly Val Glu Thr Thr Phe Tyr Tyr Tyr Tyr 115 120 125 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 140 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 145 150 155 160 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 165 170 175 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 180 185 190 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 195 200 205 Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr 210 215 220 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 225 230 235 240 Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro 245 250 255 Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 275 280 285 Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly 290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn 305 310 315 320 Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro 340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 405 410 415 Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 450 455 460 Ser Leu Ser Pro Gly Lys 465 470 31470PRTArtificial Sequenceheavy chain immunoglobulin 31Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 85 90 95 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Lys Gly Tyr Ser Ser Gly Trp Tyr Tyr Tyr Tyr Tyr 115 120 125 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 140 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 145 150 155 160 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 165 170 175 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 180 185 190 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 195 200 205 Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr 210 215 220 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 225 230 235 240 Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro 245 250 255 Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 275 280 285 Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly 290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn 305 310 315 320 Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro 340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 405 410 415 Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 450 455 460 Ser Leu Ser Pro Gly Lys 465 470 32470PRTArtificial Sequenceheavy chain immunoglobulin 32Met Glu Phe Gly Leu Ser Trp Leu Phe Leu Val Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe 35 40 45 Ser Ser Tyr Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Thr Thr Phe Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Thr 85 90 95 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Lys Asp Leu Gly Trp Ser Asp Ser Tyr Tyr Tyr Tyr 115 120 125 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 140 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 145 150 155 160 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 165 170 175 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 180 185 190 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 195 200 205 Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr 210 215 220 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 225 230 235 240 Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro 245 250 255 Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 275 280 285 Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly 290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn 305 310 315 320 Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro 340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 405 410 415 Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 450 455 460 Ser Leu Ser Pro Gly Lys 465 470 33470PRTArtificial Sequenceimmunoglobulin heavy chain of 2.12.1 fx 33Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Ile Ile Lys Gly 1 5 10 15 Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys 20 25 30 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Ser Asp Tyr Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser Thr Arg Asp Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 85 90 95 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Asp Gly Val Glu Thr Thr Phe Tyr Tyr Tyr Tyr 115 120 125 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 140 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 145 150 155 160 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 165 170 175 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 180 185 190 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 195 200 205 Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr 210 215 220 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 225 230 235 240 Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro 245 250 255 Pro Val Ala

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 260 265 270 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 275 280 285 Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly 290 295 300 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn 305 310 315 320 Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp 325 330 335 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro 340 345 350 Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 355 360 365 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 370 375 380 Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 385 390 395 400 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 405 410 415 Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 420 425 430 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 435 440 445 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 450 455 460 Ser Leu Ser Pro Gly Lys 465 470 34125PRTArtificial Sequencemature immunoglobulin heavy chain variable region of 2.12.1 fx 34Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Tyr Ile Ser Ser Ser Gly Ser Thr Arg Asp Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Gly Val Glu Thr Thr Phe Tyr Tyr Tyr Tyr Tyr Gly Met 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 35236PRTArtificial Sequenceimmunoglobulin light chain of 2.12.1 fx 35Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Asp Ile Arg Arg Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Arg Leu Gln Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110 His Asn Asn Tyr Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 36108PRTArtificial Sequencemature immunoglobulin light chain variable region of 2.12.1 fx 36Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg Arg Asp 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Arg Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Asn Tyr Pro Arg 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 37112PRTArtificial Sequencehumanized 7C10 immunoglobulin light chain variable region; version 1 37Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Gln Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Leu Tyr Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 38112PRTArtificial Sequencehumanized 7C10 immunoglobulin light chain variable region; version 2 38Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Gln Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Leu Tyr Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 39117PRTArtificial Sequencehumanized 7C10 immunoglobulin heavy chain variable region; version 1 39Gln Val Gln Leu Gln 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 Tyr Ser Ile Thr Gly Gly 20 25 30 Tyr Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu 50 55 60 Lys Asp Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly Arg Val Phe Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 40117PRTArtificial Sequencehumanized 7C10 immunoglobulin heavy chain variable region; version 2 40Gln Val Gln Leu Gln 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 Tyr Ser Ile Thr Gly Gly 20 25 30 Tyr Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu 50 55 60 Lys Asp Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly Arg Val Phe Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 41117PRTArtificial Sequencehumanized 7C10 immunoglobulin heavy chain variable region; version 3 41Gln Val Gln Leu Gln 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 Tyr Ser Ile Ser Gly Gly 20 25 30 Tyr Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu 50 55 60 Lys Asp Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Gly Arg Val Phe Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 42130PRTArtificial SequenceA12 immunoglobulin heavy chain variable region 42Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ala Pro Leu Arg Phe Leu Glu Trp Ser Thr Gln Asp His Tyr 100 105 110 Tyr Tyr Tyr Tyr Met Asp Val Trp Gly Lys Gly Thr Thr Val Thr Val 115 120 125 Ser Ser 130 43109PRTArtificial SequenceA12 immunoglobulin light chain variable region 43Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 1 5 10 15 Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Asn Ser Asp Asn Arg 85 90 95 Leu Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ser 100 105 44119PRTArtificial Sequence1A immunoglobulin heavy chain variable region 44Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Arg Asn Tyr 20 25 30 Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Gly Ser Gly Gly Gly Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Ala Pro Asn Trp Gly Ser Asp Ala Phe Asp Ile Trp Gly Gln Gly 100 105 110 Thr Met Val Thr Val Ser Ser 115 45107PRTArtificial Sequence1A immunoglobulin light chain variable region 45Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Pro 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 46251PRTArtificial Sequencesingle chain fv 8A1 46Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Thr Ile Ser Cys Lys Gly Pro Gly Tyr Asn Phe Phe Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Tyr Pro Thr Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Val Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Ser Ile Arg Tyr Cys Pro Gly Gly Arg Cys Tyr Ser Gly Tyr 100 105 110 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 125 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser 130 135 140 Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val 145 150 155 160 Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp 165 170 175 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys 180 185 190 Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 195 200 205 Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu 210 215 220 Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val Val 225 230 235 240 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 245 250 47245PRTArtificial Sequencesingle chain fv 9A2 47Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Arg Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Thr Ser Gly Tyr Thr Phe Arg Asn Tyr 20 25 30 Asp Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Ser Gly His Tyr Gly Asn Thr Asp His Ala Gln Lys Phe 50 55 60 Gln Gly Arg Phe Thr Met Thr Lys Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Phe Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gln Trp Asn Val Asp Tyr Trp Gly Arg Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Ala Leu Asn Phe Met Leu Thr Gln Pro His Ser Val Ser 130 135 140 Glu Ser Pro Gly Lys Thr Val Thr Ile Ser Cys Thr Arg Ser Ser Gly 145 150 155 160 Ser Ile Ala Ser Asn Tyr Val Gln Trp Tyr Gln Gln Arg Pro Gly Ser 165

170 175 Ser Pro Thr Thr Val Ile Phe Glu Asp Asn Arg Arg Pro Ser Gly Val 180 185 190 Pro Asp Arg Phe Ser Gly Ser Ile Asp Thr Ser Ser Asn Ser Ala Ser 195 200 205 Leu Thr Ile Ser Gly Leu Lys Thr Glu Asp Glu Ala Asp Tyr Tyr Cys 210 215 220 Gln Ser Phe Asp Ser Thr Asn Leu Val Val Phe Gly Gly Gly Thr Lys 225 230 235 240 Val Thr Val Leu Gly 245 48245PRTArtificial Sequencesingle chain fv 11A4 48Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ser Ser Pro Tyr Ser Ser Arg Trp Tyr Ser Phe Asp Pro Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Ser Tyr Glu Leu Thr Gln 130 135 140 Pro Pro Ser Val Ser Val Ser Pro Gly Gln Thr Ala Thr Ile Thr Cys 145 150 155 160 Ser Gly Asp Asp Leu Gly Asn Lys Tyr Val Ser Trp Tyr Gln Gln Lys 165 170 175 Pro Gly Gln Ser Pro Val Leu Val Ile Tyr Gln Asp Thr Lys Arg Pro 180 185 190 Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser Asn Ser Gly Asn Ile Ala 195 200 205 Thr Leu Thr Ile Ser Gly Thr Gln Ala Val Asp Glu Ala Asp Tyr Tyr 210 215 220 Cys Gln Val Trp Asp Thr Gly Thr Val Val Phe Gly Gly Gly Thr Lys 225 230 235 240 Leu Thr Val Leu Gly 245 49251PRTArtificial Sequencesingle chain fv 7A4 49Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Thr Ile Ser Cys Lys Gly Ser Gly Tyr Asn Phe Phe Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Asp Leu Glu Trp Met 35 40 45 Gly Ile Ile Tyr Pro Thr Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Val Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Ser Ile Arg Tyr Cys Pro Gly Gly Arg Cys Tyr Ser Gly Tyr 100 105 110 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 125 Gly Gly Gly Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser 130 135 140 Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val 145 150 155 160 Arg Ile Thr Cys Arg Gly Asp Ser Leu Arg Asn Tyr Tyr Ala Ser Trp 165 170 175 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys 180 185 190 Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 195 200 205 Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu 210 215 220 Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Met Val 225 230 235 240 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 245 250 50249PRTArtificial Sequencesingle chain fv 11A1 50Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Phe 20 25 30 Ala Met His Trp Val Arg Gln Ile Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Ser Gly Leu Arg His Asp Gly Ser Thr Ala Tyr Tyr Ala Gly Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Val Thr Gly Ser Gly Ser Ser Gly Pro His Ala Phe Pro Val Trp Gly 100 105 110 Lys Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Ala Leu Ser Tyr Val Leu Thr Gln 130 135 140 Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser Cys 145 150 155 160 Ser Gly Ser Asn Ser Asn Ile Gly Thr Tyr Thr Val Asn Trp Phe Gln 165 170 175 Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Ser Asn Asn Gln 180 185 190 Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr 195 200 205 Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala Asp 210 215 220 Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Asn Gly Pro Val Phe Gly 225 230 235 240 Gly Gly Thr Lys Val Thr Val Leu Gly 245 51251PRTArtificial Sequencesingle chain fv 7A6 51Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Thr Ile Ser Cys Lys Gly Ser Gly Tyr Asn Phe Phe Asn Tyr 20 25 30 Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 Gly Ile Ile Tyr Pro Thr Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60 Gln Gly Gln Val Thr Ile Ser Val Asp Lys Ser Ile Ser Thr Ala Tyr 65 70 75 80 Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Ser Ile Arg Tyr Cys Pro Gly Gly Arg Cys Tyr Ser Gly Tyr 100 105 110 Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser 130 135 140 Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr Val 145 150 155 160 Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Thr Asn Trp 165 170 175 Phe Gln Gln Lys Pro Gly Gln Ala Pro Leu Leu Val Val Tyr Ala Lys 180 185 190 Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 195 200 205 Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu 210 215 220 Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val Val 225 230 235 240 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 245 250 525PRTArtificial SequenceCDR 52Ser Tyr Trp Met His 1 5 5317PRTArtificial SequeceCDR 53Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Arg 5415PRTArtificial SequenceCDR 54Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp Val 1 5 10 15 5516PRTArtificial SequenceCDR 55Arg Ser Ser Gln Ser Ile Val His Ser Asn Val Asn Thr Tyr Leu Glu 1 5 10 15 567PRTArtificial SequenceCDR 56Lys Val Ser Asn Arg Phe Ser 1 5 579PRTArtificial SequenceCDR 57Phe Gln Gly Ser His Val Pro Pro Thr 1 5 58123PRTArtificial Sequenceheavy chain immunoglobulin variable region 58Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe 85 90 95 Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 115 120 59118PRTArtificial Sequenceheavy chain immunoglobulin variable region 59Gln Val Gln Phe Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Leu Met His Trp Ile Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Asn Asn Val Val Thr Lys Phe Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Pro Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Ala Tyr Cys Arg Pro Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser 115 60123PRTArtificial Sequenceheavy chain immunoglobulin variable region 60Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Arg Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe 85 90 95 Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp 100 105 110 Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser 115 120 61120PRTArtificial Sequenceheavy chain immunoglobulin variable region 61Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Ser Phe 20 25 30 Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Leu Pro Gly Ser Gly Gly Thr His Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Phe Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly His Ser Tyr Tyr Phe Tyr Asp Gly Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 62120PRTArtificial Sequenceheavy chain immunoglobulin variable region 62Gln Val Gln Leu Gln Gln Pro Gly Ser Val Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Ser 20 25 30 Trp Ile His Trp Ala Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile His Pro Asn Ser Gly Asn Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Val Asp Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Arg Tyr Gly Ser Pro Tyr Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Leu Thr Val Ser Ser 115 120 63120PRTArtificial Sequenceheavy chain immunoglobulin variable region 63Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Asn Ser Gly Gly Thr Lys Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Pro Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Asp Tyr Tyr Gly Ser Ser Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Leu Thr Val Ser Ser 115 120 64123PRTArtificial Sequenceheavy chain immunoglobulin variable region 64Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe 85 90 95 Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 115 120 65124PRTArtificial Sequenceheavy chain immunoglobulin variable region 65Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Arg Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe 85 90 95 Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp 100 105 110 Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 115 120 66124PRTArtificial Sequenceheavy chain immunoglobulin variable region 66Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe 50 55 60

Gln Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe 85 90 95 Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 67120PRTArtificial Sequenceheavy chain immunoglobulin variable region 67Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Asn Ser Gly Gly Thr Lys Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Pro Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Tyr Asp Tyr Tyr Gly Ser Ser Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 68117PRTArtificial Sequenceheavy chain immunoglobulin variable region 68Gln Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Ile His Trp Val Lys Gln Arg Pro Gly Glu Gly Leu Glu Trp Ile 35 40 45 Gly Trp Ile Tyr Pro Gly Ser Gly Asn Thr Lys Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Gly Gly Lys Phe Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser 100 105 110 Val Thr Val Ser Ser 115 69124PRTArtificial Sequenceheavy chain immunoglobulin variable region 69Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Arg Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe 85 90 95 Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp 100 105 110 Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser 115 120 70120PRTArtificial Sequenceheavy chain immunoglobulin variable region 70Gln Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Ile Asn Trp Met Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Trp Ile Asp Pro Gly Ser Gly Asn Thr Lys Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Arg Glu Lys Thr Thr Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ala 115 120 71115PRTArtificial Sequenceheavy chain immunoglobulin variable region 71Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Met Lys Pro Gly Ala Ser 1 5 10 15 Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asp Tyr Trp 20 25 30 Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile Gly 35 40 45 Glu Ile Leu Pro Gly Ser Gly Ser Thr Asn Tyr His Glu Arg Phe Lys 50 55 60 Gly Lys Ala Thr Phe Thr Ala Asp Thr Ser Ser Ser Thr Ala Tyr Met 65 70 75 80 Gln Leu Asn Ser Leu Thr Ser Glu Asp Ser Gly Val Tyr Tyr Cys Leu 85 90 95 His Gly Asn Tyr Asp Phe Asp Gly Trp Gly Gln Gly Thr Thr Leu Thr 100 105 110 Val Ser Ser 115 72120PRTArtificial Sequenceheavy chain immunoglobulin variable region 72Gln Val Gln Leu Leu Glu Ser Gly Ala Glu Leu Met Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Ser Ser Phe 20 25 30 Trp Ile Glu Trp Val Lys Gln Arg Pro Gly His Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Leu Pro Gly Ser Gly Gly Thr His Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Phe Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly His Ser Tyr Tyr Phe Tyr Asp Gly Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 73113PRTArtificial Sequencelight chain immunoglobulin variable region 73Asp Val Leu Met Thr Gln Ile Pro Val Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ile Ile Val His Asn 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 74113PRTArtificial Sequencelight chain immunoglobulin variable region 74Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 75113PRTArtificial Sequencelight chain immunoglobulin variable region 75Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 76113PRTArtificial Sequencelight chain immunoglobulin variable region 76Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 77113PRTArtificial Sequencelight chain immunoglobulin variable region 77Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 78113PRTArtificial Sequencelight chain immunoglobulin variable region 78Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Xaa Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Xaa Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 79113PRTArtificial Sequencelight chain immunoglobulin variable region 79Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 80113PRTArtificial Sequencelight chain immunoglobulin variable region 80Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 81113PRTArtificial Sequencelight chain immunoglobulin variable region 81Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 82113PRTArtificial Sequencelight chain immunoglobulin variable region 82Asp Val Leu Met Thr Gln Ile Pro Val Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ile Ile Val His Asn 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 83113PRTArtificial Sequencelight chain immunoglobulin variable region 83Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Phe Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Ser Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 84113PRTArtificial Sequencelight chain immunoglobulin variable region 84Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 85113PRTArtificial

Sequencelight chain immunoglobulin variable region 85Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 86113PRTArtificial Sequencelight chain immunoglobulin variable region 86Glu Leu Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Thr Ile Val His Ser 20 25 30 Asn Gly Asp Thr Tyr Leu Asp Trp Phe Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 87113PRTArtificial Sequencelight chain immunoglobulin variable region 87Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 88113PRTArtificial Sequencelight chain immunoglobulin variable region 88Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 89113PRTArtificial Sequencelight chain immunoglobulin variable region 89Asp Val Leu Met Thr Gln Thr Pro Val Ser Leu Ser Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Thr Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Ile Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Ala 85 90 95 Ser His Ala Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 90113PRTArtificial Sequencelight chain immunoglobulin variable region 90Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Ile Val His Ser 20 25 30 Ser Gly Asn Thr Tyr Phe Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Ile Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 91113PRTArtificial Sequencelight chain immunoglobulin variable region 91Asp Ile Glu Leu Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 92113PRTArtificial Sequencelight chain immunoglobulin variable region 92Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 93113PRTArtificial Sequencelight chain immunoglobulin variable region 93Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 94113PRTArtificial Sequencelight chain immunoglobulin variable region 94Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 95113PRTArtificial Sequencelight chain immunoglobulin variable region 95Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Val Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 96113PRTArtificial Sequencelight chain immunoglobulin variable region 96Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Asn Gln Thr Ile Leu Leu Ser 20 25 30 Asp Gly Asp Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 97113PRTArtificial Sequencelight chain immunoglobulin variable region 97Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Thr Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Thr Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Thr His Ala Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 98113PRTArtificial Sequencelight chain immunoglobulin variable region 98Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Ser Ile Ser Ser Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Gln Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg 9914DNAArtificial Sequenceprimer 99ctccgcttcc tttc 1410018DNAArtificial Sequenceanti-sense 100atctctccgc ttcctttc 1810118DNAArtificial Sequenceanti-sense 101atctctccgc ttcctttc 1810219DNAArtificial Sequenceprimer 102aggagctcga ggcgttcag 1910322DNAArtificial Sequenceprimer 103gtcttgggtg ggtagagcaa tc 2210421DNAArtificial Sequenceprimer 104aggccaaacg tcaccgtccc c 21105109PRTArtificial SequenceA12 immunoglobulin light chain variable region 105Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 1 5 10 15 Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Thr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Ile Leu Val Ile Tyr 35 40 45 Gly Glu Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Lys Ser Arg Asp Gly Ser Gly Gln His 85 90 95 Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly 100 105 10611PRTArtificial SequenceCDR-L1 106Arg Ala Ser Gln Ser Ile Gly Ser Ser Leu His 1 5 10 1077PRTArtificial SequenceCDR-L2' 107Tyr Ala Ser Gln Ser Leu Ser 1 5 1089PRTArtificial SequenceCDR-L3 108His Gln Ser Ser Arg Leu Pro His Thr 1 5 1095PRTArtificial SequenceCDR-H1 109Ser Phe Ala Met His 1 5 11010PRTArtificial SequenceCDR-H1 alternate 110Gly Phe Thr Phe Ser Ser Phe Ala Met His 1 5 10 11116PRTArtificial SequenceCDR-H2 111Val Ile Asp Thr Arg Gly Ala Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 11210PRTArtificial SequenceCDR-H3 112Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val 1 5 10

Claims

1. A method for treating or preventing an APC-mediated medical disorder, in a subject, comprising administering, to the subject, a therapeutically effective amount of one or more IGF1R inhibitors or a pharmaceutical composition thereof.

2. The method of claim 1 wherein the disorder is a member selected from the group consisting of familial adenomatous polyposis (FAP), Gardner syndrome, Turcot syndrome and attenuated familial adenomatous polyposis.

3. The method of claim 1 wherein an IGF1R inhibitor is one or more members selected from the group consisting of an isolated antibody or antigen-binding fragment thereof that binds specifically to IGF1R, ##STR00017##

4. The method of claim 3 wherein the antibody or antigen-binding fragment thereof comprises one or more CDRs from a light chain variable region comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 37 or 38 and/or one or more CDRs from a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, 12, 39, 40 or 41.

5. The method of claim 4 wherein the antibody or fragment comprises a light chain immunoglobulin amino acid sequence which comprises CDR-L1 comprising amino acids RASQSIGSSLH (SEQ ID NO: 106), CDR-L2 comprising amino acids YASQSLS (SEQ ID NO: 107) and CDR-L3 comprising amino acids HQSSRLPHT (SEQ ID NO: 108); and/or a heavy chain immunoglobulin amino acid sequence which comprises CDR-H1 comprising amino acids SFAMH (SEQ ID NO: 109) or GFTFSSFAMH (SEQ ID NO: 110), CDR-H2 comprising amino acids VIDTRGATYYADSVKG (SEQ ID NO: 111) and CDR-H3 comprising amino acids LGNFYYGMDV (SEQ ID NO: 112).

6. The method of claim 5 wherein the antibody is an isolated antibody comprising a light chain variable region comprising amino acids 20-128 of SEQ ID NO: 2, 4, 6 or 8 and/or a heavy chain variable region comprising amino acids 20-137 of SEQ ID NO: 10 or 12.

7. The method of claim 3 wherein the antibody or fragment comprises a light chain immunoglobulin variable region linked to a kappa light chain constant region.

8. The method of claim 3 wherein the antibody or fragment comprises a heavy chain immunoglobulin variable region linked to a gamma 1 heavy chain constant region.

9. The method of claim 3 wherein the antibody or fragment comprises a mature heavy chain immunoglobulin encoded by a polynucleotide in plasmid 15H12/19D12 HCA (γ4) which is obtainable from a cell line deposited at the American Type Culture Collection (ATCC) under number PTA-5214; and/or a mature light chain immunoglobulin encoded by a polynucleotide in plasmid 15H12/19D12 LCF (κ) which is obtainable from a cell line deposited at the American Type Culture Collection (ATCC) under number PTA-5220.

10. The method of claim 1 wherein an inhibitor is administered to the subject in association with one or more therapeutic procedures, diagnostic procedures or additional chemotherapeutic agents.

11. The method of claim 10 wherein an additional chemotherapeutic agent is one or more non-steroidal anti-inflammatory agents.

12. The method of claim 10 wherein an additional chemotherapeutic agent is one or members selected from the group consisting of piroxicam, sulindac, aptosyn (sulindac sulfone), indomethacin, rofecoxib, celecoxib, aspirin, a dietary calcium supplement, a retinoid, a carotenoid, ascorbic acid, α-tocopherol, selenium, folate and methionine.

13. The method of claim 1 wherein an IGF1R inhibitor and an additional chemotherapeutic agent are in a single pharmaceutical composition along with a pharmaceutically acceptable carrier.

14. The method of claim 1 wherein an IGF1R inhibitor and an additional chemotherapeutic agent are in two or more separate pharmaceutical compositions each along with pharmaceutically acceptable carriers.

15. The method of claim 1 wherein an inhibitor is administered to the subject by a parenteral route.

16. The method of claim 3 wherein the antibody or antigen-binding fragment thereof is a member selected from the group consisting of a monoclonal antibody, a polyclonal antibody, an anti-idiotypic antibody, a chimeric antibody, a bispecific antibody, a humanized antibody, a fully human antibody, a recombinant antibody, a Fab, a F(ab)₂, an Fv, a single chain antibody, a dsFv and a linear antibody.

17. The method of claim 1 wherein the subject is human.

18. A composition comprising one or more IGF1R inhibitors in association with one or more non-steroidal anti-inflammatory agents or a pharmaceutical composition thereof.

19. The composition of claim 18 wherein the agent is one or more members selected from the group consisting of piroxicam, sulindac, aptosyn (sulindac sulfone), indomethacin, rofecoxib, celecoxib, aspirin, a dietary calcium supplement, a retinoid, a carotenoid, ascorbic acid, α-tocopherol, selenium, folate and methionine.

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