IL-18 binding protein (IL-18BP) and antibodies in inflammatory diseases

Abstract

The present invention provides means and methods for treating Interleukin 18 (IL-18)-associated diseases and disorders. In particular, the present invention discloses antibodies specific for free IL-18 and IL-18 Binding Protein (IL-18BP) for use in such treatments and for the diagnosis of the diseases and disorders.

Claims

1. A composition comprising a pharmaceutically acceptable carrier and a recombinant IL-18 binding protein (IL-18BP) having the amino acid sequence of SEQ ID NO: 7, and at least one deletion variant comprising a deletion of 1 to 5 amino acid residues at the C-terminal end of the IL-18BP, wherein the proportion of the at least one deletion variant in the composition is less than 30%.

2. The composition of claim 1, wherein the proportion of the at least one deletion variant in the composition is less than 20%.

3. The composition of claim 2, wherein the proportion of the at least one deletion variant in the composition is less than 15%.

4. The composition of claim 3, wherein the proportion of the at least one deletion variant in the composition is less than 10%.

5. The composition of claim 4, wherein the proportion of the at least one deletion variant in the composition is less than 7.5%.

6. The composition of claim 5, wherein the proportion of the at least one deletion variant in the composition is less than 5%.

7. The composition of claim 6, wherein the proportion of the at least one deletion variant in the composition is less than 2.5%.

8. The composition of claim 7, wherein the proportion of the at least one deletion variant in the composition is less than 1%.

9. The composition of claim 1, wherein the composition comprises sodium chloride, sodium dihydrogen phosphate monohydrate, disodium phosphate dehydrate, and water.

10. The composition of claim 1, wherein the recombinant IL-18BP binds to IL-18 with a binding affinity of 20-50 pM based on BIAcore measurements.

11. A method of preparing the composition of claim 1, comprising growing a recombinant Chinese Hamster Ovary (CHO) cell under conditions to express the recombinant IL-18BP, and purifying the recombinant IL-18BP and the at least one deletion variant from the CHO cell.

12. The method of claim 11, wherein the recombinant IL-18BP and the at least one deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP and the at least one deletion variant; (c) capturing the recombinant IL-18BP and the at least one deletion variant on an anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP and the at least one deletion variant; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP and the at least one deletion variant.

13. A composition comprising a pharmaceutically acceptable carrier and a recombinant IL-18 binding protein (IL-18BP) having the amino acid sequence of SEQ ID NO: 7, and at least one deletion variant comprising a deletion of 1 to 30 amino acid residues at the N-terminal end of the IL-18BP, wherein the proportion of the at least one deletion variant in the composition is less than 30%.

14. The composition of claim 13, wherein the proportion of the at least one deletion variant in the composition is less than 20%.

15. The composition of claim 14, wherein the proportion of the at least one deletion variant in the composition is less than 15%.

16. The composition of claim 15, wherein the proportion of the at least one deletion variant in the composition is less than 10%.

17. The composition of claim 16, wherein the proportion of the at least one deletion variant in the composition is less than 7.5%.

18. The composition of claim 17, wherein the proportion of the at least one deletion variant in the composition is less than 5%.

19. The composition of claim 18, wherein the proportion of the at least one deletion variant in the composition is less than 2.5%.

20. The composition of claim 19, wherein the proportion of the at least one deletion variant in the composition is less than 1%.

21. The composition of claim 13, wherein the composition comprises sodium chloride, sodium dihydrogen phosphate monohydrate, disodium phosphate dehydrate, and water.

22. The composition of claim 13, wherein the recombinant IL-18BP binds to IL-18 with a binding affinity of 20-50 pM based on BIAcore measurements.

23. A method of preparing the composition of claim 13, comprising growing a recombinant Chinese Hamster Ovary (CHO) cell under conditions to express the recombinant IL-18BP, and purifying the recombinant IL-18BP and the at least one deletion variant from the CHO cell.

24. The method of claim 23, wherein the recombinant IL-18BP and the at least one deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP and the at least one deletion variant; (c) capturing the recombinant IL-18BP and the at least one deletion variant on an anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP and the at least one deletion variant; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP and the at least one deletion variant.

25. A composition comprising a pharmaceutically acceptable carrier and a recombinant IL-18 binding protein (IL-18BP) having the amino acid sequence of SEQ ID NO: 7, at least one first deletion variant comprising a deletion of 1 to 5 amino acid residues at the C-terminal end of the IL-18BP, and at least one second deletion variant comprising a deletion of 1 to 30 amino acid residues at the N-terminal end of the IL-18BP, wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 30%.

26. The composition of claim 25, wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 20%.

27. The composition of claim 26, wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 15%.

28. The composition of claim 27, wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 10%.

29. The composition of claim 28, wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 7.5%.

30. The composition of claim 29, wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 5%.

31. The composition of claim 25, wherein the composition comprises sodium chloride, sodium dihydrogen phosphate monohydrate, disodium phosphate dehydrate, and water.

32. The composition of claim 25, wherein the recombinant IL-18BP binds to IL-18 with a binding affinity of 20-50 pM based on BIAcore measurements.

33. A method of preparing the composition of claim 25, comprising growing a recombinant Chinese Hamster Ovary (CHO) cell under conditions to express the recombinant IL-18BP, and purifying the recombinant IL-18BP, the at least one first deletion variant and the at least one second deletion variant from the CHO cell.

34. The method of claim 33, wherein the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant; (c) capturing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant on an anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP, the at least one first deletion variant and the at least one second deletion variant.

35. The method of claim 34, wherein the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant and diafiltrating against a basic borate buffer at a pH higher than pH 7.5; (c) capturing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant on a TMAE Hi-Cap anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant with a borate buffer containing NaCl; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1: Comparison of total and free IL-18 in individual sepsis patients. Adapted from Novick et al 2001. The level of free IL-18 (closed circles) in sera of sepsis patients upon admission was calculated based on the concentration of total IL-18 (open circles) and IL-18BPa, taking into account a 1:1 complex of IL-18 and IL-18BPa and a calculated KD of 400 pM. Each vertical line links total and free IL-18 in an individual serum sample. The above ELISA assays are performed with the pair of antibodies developed by Taniguchi et al 1997 1, namely antibodies 125-2H as primary/capture antibody and 159-12B as secondary/developing antibody.

(2) FIG. 2: Detection of total IL-18 with antibodies 125-2H and 159-12B. The data indicates that both antibodies quantify total IL-18.

(3) FIG. 3: Titration of 400 pg/ml IL-18 as a function of IL-18BP level

(4) FIG. 4: Mouse IL-18 induction in the lung airway space at day 5 after 1) air exposure, 2) tobacco smoke (TS), 3) p[I:C] alone, 4) p[I:C] combined to tobacco smoke at day 4 (induction of exacerbation). Dotted line indicates lower limit of detection. Statistical analyses were performed using either the unpaired t-test.

(5) FIG. 5: Inhibition of total cell infiltration in the mouse lung airway space at day 5 after 1) air exposure, 2) tobacco smoke (TS), 3) p[I:C] alone, 4) p[I:C] combined to tobacco smoke at day 4 (induction of exacerbation), 5-7) p[I:C] combined to tobacco smoke at day 4 under IL-18BP treatment at either 1, 3 or 10 mg/kg, 8) dexamethasone treatment at 10 mg/kg. Statistical analyses were performed using either the unpaired t-test.

(6) FIG. 6: Inhibition of total cell infiltration in the mouse lung airway space at day 5 after 1) air exposure, 2) tobacco smoke (TS), 3) p[I:C] alone, 4) p[I:C] combined to tobacco smoke at day 4 (induction of exacerbation), 5) p[I:C] combined to tobacco smoke at day 4 under IL-18BP treatment at 10 mg/kg, 6) dexamethasone treatment at 10 mg/kg. Statistical analyses were performed using ANOVA test (post-test Sidak's).

(7) FIG. 7: Inhibition of neutrophil infiltration by IL-18BP. Neutrophil infiltration in the mouse lung airway space was monitored at day 5 after 1) air exposure, 2) tobacco smoke (TS), 3) p[I:C] alone, 4) p[I:C] combined to tobacco smoke at day 4 (induction of exacerbation), 5-7) p[I:C] combined to tobacco smoke at day 4 under IL-18BP treatment at either 1, 3 or 10 mg/kg, 8) dexamethasone treatment at 10 mg/kg. Statistical analyses were performed using ANOVA test (post-test Sidak's).

(8) FIG. 8: Inhibition of neutrophil infiltration by IL-18BP. Neutrophil infiltration in the mouse lung airway space was monitored at day 5 after 1) air exposure, 2) tobacco smoke (TS), 3) p[I:C] alone, 4) p[I:C] combined to tobacco smoke at day 4 (induction of exacerbation), 5) p[I:C] combined to tobacco smoke at day 4 under IL-18BP treatment at 10 mg/kg, 6) dexamethasone treatment at 10 mg/kg. Statistical analyses were performed using ANOVA test (post-test Sidak's).

(9) FIG. 9: Inhibition of G-CSF pathway by IL-18BP. The presence of G-CSF (pg/ml) was monitored in the mouse lung airway space by ELISA at day 5 after 1) air exposure, 2) tobacco smoke (TS), 3) p[I:C] alone, 4) p[I:C] combined to tobacco smoke at day 4 (induction of exacerbation), 5-7) p[I:C] combined to tobacco smoke at day 4 under IL-18BP treatment at either 1, 3 or 10 mg/kg, 8) dexamethasone treatment at 10 mg/kg. Dotted line indicates lower limit of detection. Statistical analyses were performed using Students t-test.

(10) FIG. 10: Mitigation of weight loss by IL-18BP. Mouse weight loss was monitored at day 5 after 1) air exposure, 2) tobacco smoke (TS), 3) p[I:C] alone, 4) p[I:C] combined to tobacco smoke at day 4 (induction of exacerbation), 5) p[I:C] combined to tobacco smoke at day 4 under IL-18BP treatment at 10 mg/kg, 7) dexamethasone treatment at 10 mg/kg. Statistical analyses were performed using ANOVA test (post-test Sidak's).

(11) FIG. 11: shows the amino acid sequences of the variable heavy chain (VH) and the variable light chain (VK) of antibody 131B4-2. The complementary determining regions CDR 1, CDR 2 and CDR 3 are identified by underlining the respective sequences as determined by the Kabat numbering system. From left to right, the first underlined bold sequence in each of the VH and VK sequences shown represents CDR1, the second underlined bold sequence represents CDR 2 and the third underlined bold sequence represents CDR 3.

(12) FIG. 12: shows the amino acid sequences of isoforms a, b, c and d of the IL-18BP

SEQUENCES

(13) SEQ ID NO 1: IL-18 Epitope 1: Tyr-Phe-Gly-Lys-Leu-Glu-Ser-Lys-Leu-Ser-Val-Ile-Arg-Asn

(14) SEQ ID NO 2: IL-18 Epitope 2: Phe-Ile-Ile-Ser-Met-Tyr-Lys-Asp-Ser-Gln-Pro-Arg-Gly-Met-Ala-Val-Thre-Ile-Ser-Val-Lys

(15) SEQ ID NO 3: IL-18 Epitope 3: Glu-Met-Asn-Pro-Pro-Asp-Asn-Ile-Lys-Asp-Thr-Lys-Ser-Asp-Ile-Ile-Phe

(16) SEQ ID NO 4: IL-18 Epitope 4: Tyr-Phe-Gly-Lys-Leu-Glu-Ser

(17) SEQ ID NO 5: IL-18 Epitope 5: Tyr-Lys-Asp-Ser-Gln-Pro-Arg-Gly-Met-Ala

(18) SEQ ID NO 6: IL-18 Epitope 6: Asp-Asn-Ile-Lys-Asp-Thr-Lys

(19) SEQ ID NO 7: IL-18 Binding Protein (IL-18BP)

(20) SEQ ID NO 8: 13-amino acid Linker Sequence: Glu-Phe-Gly-Aa-Gly-Leu-Val-Leu-Gly-Gly-Gln-Phe-Met

(21) SEQ ID NO 9: Antibody 107C6 VH variable domain sequence

(22) SEQ ID NO 10: Antibody 107C6 VK variable domain sequence

(23) SEQ ID NO 11: Antibody 108F8 VH variable domain sequence

(24) SEQ ID NO 12: Antibody 108F8 VK variable domain sequence

(25) SEQ ID NO 13: Antibody 109A6 VH variable domain sequence

(26) SEQ ID NO 14: Antibody 109A6 VK variable domain sequence

(27) SEQ ID NO 15: Antibody 111A6 VH variable domain sequence

(28) SEQ ID NO 16: Antibody 111A6 VK variable domain sequence 1

(29) SEQ ID NO 17: Antibody 111A6 VK variable domain sequence 2

(30) SEQ ID NO 18: Antibody 131B4 VH variable domain sequence

(31) SEQ ID NO 19: Antibody 131B4 and 131B4-2 VK variable domain sequence

(32) SEQ ID NO 20: Antibody 131E8 VH variable domain sequence 1

(33) SEQ ID NO 21: Antibody 131E8 VH variable domain sequence 2

(34) SEQ ID NO 22: Antibody 131E8 VK variable domain sequence

(35) SEQ ID NO 23: Antibody 132H4 VH variable domain sequence

(36) SEQ ID NO 24: Antibody 132H4 VK variable domain sequence

(37) SEQ ID NO 25: Antibody 133A6 VH variable domain sequence

(38) SEQ ID NO 26: Antibody 133A6 VK variable domain sequence

(39) SEQ ID NO 27: Antibody 107C6 VH sequence CDR1: Gly Tyr Thr Phe Thr Asn Tyr Gly

(40) SEQ ID NO 28: Antibody 107C6 VH sequence CDR2; Ile Asn Thr Tyr Ser Gly Val Pro

(41) SEQ ID NO 29: Antibody 107C6 VH sequence CDR3: Ala Arg Glu Gly Tyr Ser Thr Thr Arg Ser Met Asp Tyr

(42) SEQ ID NO 30: Antibody 107C6 VK sequence CDR1: Gln Ser Leu Leu Asp Ser Arg Thr Arg Lys Asn Tyr

(43) SEQ ID NO 31: Antibody 107C6 VK sequence CDR2: Trp Ala Ser

(44) SEQ ID NO 32: Antibody 107C6 VK sequence CDR3: Lys Gln Ser Tyr Asn Leu Arg Thr

(45) SEQ ID NO 33: Antibody 108F8 VH sequence CDR1: Gly Tyr Thr Phe Thr Asn Tyr Gly

(46) SEQ ID NO 34: Antibody 108F8 VH sequence CDR2: Ile Asn Thr Tyr Ser Gly Val Pro

(47) SEQ ID NO 35: Antibody 108F8 VH sequence CDR3: Ala Arg Glu Gly Tyr Ser Thr Thr Arg Ser Met Asp Tyr

(48) SEQ ID NO 36: Antibody 108F8 VK sequence CDR1: Gln Ser Leu Leu Asp Ser Arg Thr Arg Lys Asn Tyr

(49) SEQ ID NO 37: Antibody 108F8 VK sequence CDR2: Trp Ala Ser

(50) SEQ ID NO 38: Antibody 108F8 VK sequence CDR3: Lys Gln Ser Tyr Asn Leu Arg Thr

(51) SEQ ID NO 39: Antibody 109A6 VH sequence CDR1: Gly Phe Lys Ile Lys Asp Thr Tyr

(52) SEQ ID NO 40: Antibody 109A6 VH sequence CDR2: Ile Asp Pro Ala Asn Gly Asn Thr

(53) SEQ ID NO 41: Antibody 109A6 VH sequence CDR3: Ala Gly Tyr Val Trp Phe Ala Tyr

(54) SEQ ID NO 42: Antibody 109A6 VK sequence CDR1: Gln Arg Leu Val His Ser Asn Gly Asn Thr Tyr

(55) SEQ ID NO 43: Antibody 109A6 VK sequence CDR2: Thr Val Ser

(56) SEQ ID NO 44: Antibody 109A6 VK sequence CDR3: Ser Gln Ser Thr Leu Val Pro Trp Thr

(57) SEQ ID NO 45: Antibody 111A6 VH sequence CDR1: Gly Phe Lys Ile Lys Asp Thr Tyr

(58) SEQ ID NO 46: Antibody 111A6 VH sequence CDR2: Ile Asp Pro Ala Asn Gly Asn Thr

(59) SEQ ID NO 47: Antibody 111A6 VH sequence CDR3: Ala Gly Tyr Val Trp Phe Ala Tyr

(60) SEQ ID NO 48: Antibody 111A6 VK sequence 1 CDR1: Ser Ser Val Ser Ser Ser Tyr

(61) SEQ ID NO 49: Antibody 111A6 VK sequence 1 CDR2: Ser Thr Ser

(62) SEQ ID NO 50: Antibody 111A6 VK sequence 1 CDR3: Gln Gln Tyr Ser Gly Tyr Pro Leu Thr

(63) SEQ ID NO 51: Antibody 111A6 VK sequence 2 CDR1: Gln Arg Leu Val His Ser Asn Gly Asn Thr Tyr

(64) SEQ ID NO 52: Antibody 111A6 VK sequence 2 CDR2: Thr Val Ser

(65) SEQ ID NO 53: Antibody 111A6 VK sequence 2 CDR3: Ser Gln Ser Thr Leu Val Pro Trp Thr

(66) SEQ ID NO 54: Antibody 131B4 VH sequence CDR1: Gly Phe Lys Ile Lys Asp Thr Tyr

(67) SEQ ID NO 55: Antibody 131B4 VH sequence CDR2: Ile Asp Pro Ala Asn Gly Asn Thr

(68) SEQ ID NO 56: Antibody 131B4 VH sequence CDR3: Ala Gly Tyr Val Trp Phe Ala Tyr

(69) SEQ ID NO 57: Antibody 131B4 VK sequence CDR1: Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr

(70) SEQ ID NO 58: Antibody 131B4 VK sequence CDR2: Lys Val Ser

(71) SEQ ID NO 59: Antibody 131B4 VK sequence CDR3: Ser Gln Ser Ser Leu Val Pro Trp Thr

(72) SEQ ID NO 60: Antibody 131E8 VH sequence 1 CDR1: Gly Phe Ser Leu Pro Asn Tyr Gly

(73) SEQ ID NO 61: Antibody 131E8 VH sequence 1 CDR2: Ile Trp Ser Gly Gly Ser Thr

(74) SEQ ID NO 62: Antibody 131E8 VH sequence 1 CDR3: Ala Arg Asn Phe Tyr Ser Lys Tyr Asp Tyr Ala Met Asp Tyr

(75) SEQ ID NO 63: Antibody 131E8 VH sequence 2 CDR1: Gly Tyr Thr Phe Thr Ser Tyr Trp

(76) SEQ ID NO 64: Antibody 131E8 VH sequence 2 CDR2: Ile Asn Pro Asn Ser Gly Ser Thr

(77) SEQ ID NO 65: Antibody 131E8 VH sequence 2 CDR3: Ala Arg Leu Gly Asp Tyr

(78) SEQ ID NO 66: Antibody 131E8 VK sequence CDR1: Ser Ser Val Ser Tyr

(79) SEQ ID NO 67: Antibody 131E8 VK sequence CDR2: Asp Thr Ser

(80) SEQ ID NO 68: Antibody 131E8 VK sequence CDR3: Phe Gln Gly Ser Gly Tyr Pro Leu Thr

(81) SEQ ID NO 69: Antibody 132H4 VH sequence CDR1: Gly Phe Thr Phe Ser Asn Tyr Ala

(82) SEQ ID NO 70: Antibody 132H4 VH sequence CDR2: Ile Ser Ser Gly Gly Ala Asn Ile

(83) SEQ ID NO 71: Antibody 132H4 VH sequence CDR3: Ala Arg Gly Asp Tyr Phe Asn His Phe Trp Phe Ala Tyr

(84) SEQ ID NO 72: Antibody 132H4 VK sequence CDR1: Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr

(85) SEQ ID NO 73: Antibody 132H4 VK sequence CDR2: Lys Val Ser

(86) SEQ ID NO 74: Antibody 132H4 VK sequence CDR3: Phe Gln Gly Ser His Val Pro Trp Thr

(87) SEQ ID NO 75: Antibody 133A6 VH sequence CDR1: Gly Phe Thr Phe Ser Asn Tyr Ala

(88) SEQ ID NO 76: Antibody 133A6 VH sequence CDR2: Ile Ser Ser Gly Gly Gly Asn Ile

(89) SEQ ID NO 77: Antibody 133A6 VH sequence CDR3

(90) SEQ ID NO 78: Antibody 133A6 VK sequence CDR1

(91) SEQ ID NO 79: Antibody 133A6 VK sequence CDR2

(92) SEQ ID NO 80: Antibody 133A6 VK sequence CDR3

(93) SEQ ID NO 81: Antibody 107C6 VH DNA sequence

(94) SEQ ID NO 82: Antibody 107C6 VH sequence

(95) SEQ ID NO 83: Antibody 107C6 VK DNA sequence

(96) SEQ ID NO 84: Antibody 107C6 VK sequence

(97) SEQ ID NO 85: Antibody 108F8 VH DNA sequence

(98) SEQ ID NO 86: Antibody 108F8 VH sequence

(99) SEQ ID NO 87: Antibody 108F8 VK DNA sequence

(100) SEQ ID NO 88: Antibody 108F8 VK sequence

(101) SEQ ID NO 89: Antibody 109A6 VH DNA sequence

(102) SEQ ID NO 90: Antibody 109A6 VH sequence

(103) SEQ ID NO 91: Antibody 109A6 VK DNA sequence

(104) SEQ ID NO 92: Antibody 109A6 VK sequence

(105) SEQ ID NO 93: Antibody 111A6 VH DNA sequence

(106) SEQ ID NO 94: Antibody 111A6 VH sequence

(107) SEQ ID NO 95: Antibody 111A6 VK DNA sequence 1

(108) SEQ ID NO 96: Antibody 111A6 VK sequence 1

(109) SEQ ID NO 97: Antibody 111A6 VK DNA sequence 2

(110) SEQ ID NO 98: Antibody 111A6 VK sequence 2

(111) SEQ ID NO 99: Antibody 131B4 VH DNA sequence 1

(112) SEQ ID NO 100: Antibody 131B4 VH sequence 1

(113) SEQ ID NO 101: Antibody 131B4 VH DNA sequence 2

(114) SEQ ID NO 102: Antibody 131B4 VH sequence 2

(115) SEQ ID NO 103: Antibody 131B4 VH variable domain sequence 2

(116) SEQ ID NO 104: Antibody 131B4 VH sequence 2 CDR1

(117) SEQ ID NO 105: Antibody 131B4 VH sequence 2 CDR2

(118) SEQ ID NO 106: Antibody 131B4 VH sequence 2 CDR3

(119) SEQ ID NO 107: Antibody 131B4 VH DNA sequence 3

(120) SEQ ID NO 108: Antibody 131B4 VH sequence 3

(121) SEQ ID NO 109: Antibody 131B4 VH variable domain sequence 3

(122) SEQ ID NO 110: Antibody 131B4 VH sequence 3 CDR1

(123) SEQ ID NO 111: Antibody 131B4 VH sequence 3 CDR2

(124) SEQ ID NO 112: Antibody 131B4 VH sequence 3 CDR3

(125) SEQ ID NO 113: Antibody 131B4 VK DNA sequence

(126) SEQ ID NO 114: Antibody 131B4 VK sequence

(127) SEQ ID NO 115: Antibody 131E8 VH DNA sequence 1

(128) SEQ ID NO 116: Antibody 131E8 VH sequence 1

(129) SEQ ID NO 117: Antibody 131E8 VH DNA sequence 2

(130) SEQ ID NO 118: Antibody 131E8 VH sequence 2

(131) SEQ ID NO 119: Antibody 131E8 VH DNA sequence 3

(132) SEQ ID NO 120: Antibody 131E8 VH sequence 3

(133) SEQ ID NO 121: Antibody 131E8 VH variable domain sequence 3

(134) SEQ ID NO 122: Antibody 131E8 VH sequence 3 CDR1

(135) SEQ ID NO 123: Antibody 131E8 VH sequence 3 CDR2

(136) SEQ ID NO 124: Antibody 131E8 VH sequence 3 CDR3

(137) SEQ ID NO 125: Antibody 131E8 VK DNA sequence

(138) SEQ ID NO 126: Antibody 131E8 VK sequence

(139) SEQ ID NO 127: Antibody 131H1 VH DNA sequence

(140) SEQ ID NO 128: Antibody 131H1 VH sequence

(141) SEQ ID NO 129: Antibody 131H1 VH variable domain sequence

(142) SEQ ID NO 130: Antibody 131H1 VH sequence CDR1

(143) SEQ ID NO 131: Antibody 131H1 VH sequence CDR2

(144) SEQ ID NO 132: Antibody 131H1 VH sequence CDR3

(145) SEQ ID NO 133: Antibody 131H1 VK DNA sequence 1

(146) SEQ ID NO 134: Antibody 131H1 VK sequence 1

(147) SEQ ID NO 135: Antibody 131H1 VK variable domain sequence 1

(148) SEQ ID NO 136: Antibody 131H1 VK sequence 1 CDR1

(149) SEQ ID NO 137: Antibody 131H1 VK sequence 1 CDR2

(150) SEQ ID NO 138: Antibody 131H1 VK sequence 1 CDR3

(151) SEQ ID NO 139: Antibody 131H1 VK DNA sequence 2

(152) SEQ ID NO 140: Antibody 131H1 VK sequence 2

(153) SEQ ID NO 141: Antibody 131H1 VK variable domain sequence 2

(154) SEQ ID NO 142: Antibody 131H1 VK sequence 2 CDR1

(155) SEQ ID NO 143: Antibody 131H1 VK sequence 2 CDR2

(156) SEQ ID NO 144: Antibody 131H1 VK sequence 2 CDR3

(157) SEQ ID NO 145: Antibody 132H4 VH DNA sequence

(158) SEQ ID NO 146: Antibody 132H4 VH sequence

(159) SEQ ID NO 147: Antibody 132H4 VK DNA sequence

(160) SEQ ID NO 148: Antibody 132H4 VK sequence

(161) SEQ ID NO 149: Antibody 133A6 VH DNA sequence

(162) SEQ ID NO 150: Antibody 133A6 VH sequence

(163) SEQ ID NO 151: Antibody 133A6 VK DNA sequence

(164) SEQ ID NO 152: Antibody 133A6 VK sequence

(165) SEQ ID NO 153: Antibody 107C6 VH sequence CDR1 according to Chothia

(166) SEQ ID NO 154: Antibody 107C6 VH sequence CDR2 according to Chothia

(167) SEQ ID NO 155: Antibody 107C6 VH sequence CDR3 according to Chothia

(168) SEQ ID NO 156: Antibody 107C6 VK sequence CDR1 according to Chothia

(169) SEQ ID NO 157: Antibody 107C6 VK sequence CDR2 according to Chothia

(170) SEQ ID NO 158: Antibody 107C6 VK sequence CDR3 according to Chothia

(171) SEQ ID NO 159: Antibody 108F8 VH sequence CDR1 according to Chothia

(172) SEQ ID NO 160: Antibody 108F8 VH sequence CDR2 according to Chothia

(173) SEQ ID NO 161: Antibody 108F8 VH sequence CDR3 according to Chothia

(174) SEQ ID NO 162: Antibody 108F8 VK sequence CDR1 according to Chothia

(175) SEQ ID NO 163: Antibody 108F8 VK sequence CDR2 according to Chothia

(176) SEQ ID NO 164: Antibody 108F8 VK sequence CDR3 according to Chothia

(177) SEQ ID NO 165: Antibody 109A6 VH sequence CDR1 according to Chothia

(178) SEQ ID NO 166: Antibody 109A6 VH sequence CDR2 according to Chothia

(179) SEQ ID NO 167: Antibody 109A6 VH sequence CDR3 according to Chothia

(180) SEQ ID NO 168: Antibody 109A6 VK sequence CDR1 according to Chothia

(181) SEQ ID NO 169: Antibody 109A6 VK sequence CDR2 according to Chothia

(182) SEQ ID NO 170: Antibody 109A6 VK sequence CDR3 according to Chothia

(183) SEQ ID NO 171: Antibody 111A6 VH sequence CDR1 according to Chothia

(184) SEQ ID NO 172: Antibody 111A6 VH sequence CDR2 according to Chothia

(185) SEQ ID NO 173: Antibody 111A6 VH sequence CDR3 according to Chothia

(186) SEQ ID NO 174: Antibody 111A6 VK sequence 1 CDR1 according to Chothia

(187) SEQ ID NO 175: Antibody 111A6 VK sequence 1 CDR2 according to Chothia

(188) SEQ ID NO 176: Antibody 111A6 VK sequence 1 CDR3 according to Chothia

(189) SEQ ID NO 177: Antibody 111A6 VK sequence 2 CDR1 according to Chothia

(190) SEQ ID NO 178: Antibody 111A6 VK sequence 2 CDR2 according to Chothia

(191) SEQ ID NO 179: Antibody 111A6 VK sequence 2 CDR3 according to Chothia

(192) SEQ ID NO 180: Antibody 131B4 VH sequence 1 CDR1 according to Chothia

(193) SEQ ID NO 181: Antibody 131B4 VH sequence 1 CDR2 according to Chothia

(194) SEQ ID NO 182: Antibody 131B4 VH sequence 1 CDR3 according to Chothia

(195) SEQ ID NO 183: Antibody 131B4 VH sequence 2 CDR1 according to Chothia

(196) SEQ ID NO 184: Antibody 131B4 VH sequence 2 CDR2 according to Chothia

(197) SEQ ID NO 185: Antibody 131B4 VH sequence 2 CDR3 according to Chothia

(198) SEQ ID NO 186: Antibody 131B4 VH sequence 3 CDR1 according to Chothia

(199) SEQ ID NO 187: Antibody 131B4 VH sequence 3 CDR2 according to Chothia

(200) SEQ ID NO 188: Antibody 131B4 VH sequence 3 CDR3 according to Chothia

(201) SEQ ID NO 189: Antibody 131B4 VK sequence CDR1 according to Chothia

(202) SEQ ID NO 190: Antibody 131B4 VK sequence CDR2 according to Chothia

(203) SEQ ID NO 191: Antibody 131B4 VK sequence CDR3 according to Chothia

(204) SEQ ID NO 192: Antibody 131E8 VH sequence 1 CDR1 according to Chothia

(205) SEQ ID NO 193: Antibody 131E8 VH sequence 1 CDR2 according to Chothia

(206) SEQ ID NO 194: Antibody 131E8 VH sequence 1 CDR3 according to Chothia

(207) SEQ ID NO 195: Antibody 131E8 VH sequence 2 CDR1 according to Chothia

(208) SEQ ID NO 196: Antibody 131E8 VH sequence 2 CDR2 according to Chothia

(209) SEQ ID NO 197: Antibody 131E8 VH sequence 2 CDR3 according to Chothia

(210) SEQ ID NO 198: Antibody 131E8 VH sequence 3 CDR1 according to Chothia

(211) SEQ ID NO 199: Antibody 131E8 VH sequence 3 CDR2 according to Chothia

(212) SEQ ID NO 200: Antibody 131E8 VH sequence 3 CDR3 according to Chothia

(213) SEQ ID NO 201: Antibody 131E8 VK sequence CDR1 according to Chothia

(214) SEQ ID NO 202: Antibody 131E8 VK sequence CDR2 according to Chothia

(215) SEQ ID NO 203: Antibody 131E8 VK sequence CDR3 according to Chothia

(216) SEQ ID NO 204: Antibody 131H1 VH sequence CDR1 according to Chothia

(217) SEQ ID NO 205: Antibody 131H1 VH sequence CDR2 according to Chothia

(218) SEQ ID NO 206: Antibody 131H1 VH sequence CDR3 according to Chothia

(219) SEQ ID NO 207: Antibody 131H1 VK sequence 1 CDR1 according to Chothia

(220) SEQ ID NO 208: Antibody 131H1 VK sequence 1 CDR2 according to Chothia

(221) SEQ ID NO 209: Antibody 131H1 VK sequence 1 CDR3 according to Chothia

(222) SEQ ID NO 210: Antibody 131H1 VK sequence 2 CDR1 according to Chothia

(223) SEQ ID NO 211: Antibody 131H1 VK sequence 2 CDR2 according to Chothia

(224) SEQ ID NO 212: Antibody 131H1 VK sequence 2 CDR3 according to Chothia

(225) SEQ ID NO 213: Antibody 132H4 VH sequence CDR1 according to Chothia

(226) SEQ ID NO 214: Antibody 132H4 VH sequence CDR2 according to Chothia

(227) SEQ ID NO 215: Antibody 132H4 VH sequence CDR3 according to Chothia

(228) SEQ ID NO 216: Antibody 132H4 VK sequence CDR1 according to Chothia

(229) SEQ ID NO 217: Antibody 132H4 VK sequence CDR2 according to Chothia

(230) SEQ ID NO 218: Antibody 132H4 VK sequence CDR3 according to Chothia

(231) SEQ ID NO 219: Antibody 133A6 VH sequence CDR1 according to Chothia

(232) SEQ ID NO 220: Antibody 133A6 VH sequence CDR2 according to Chothia

(233) SEQ ID NO 221: Antibody 133A6 VH sequence CDR3 according to Chothia

(234) SEQ ID NO 222: Antibody 133A6 VK sequence CDR1 according to Chothia

(235) SEQ ID NO 223: Antibody 133A6 VK sequence CDR2 according to Chothia

(236) SEQ ID NO 224: Antibody 133A6 VK sequence CDR3 according to Chothia

(237) SEQ ID NO 225: Antibody 131B4-2 VH sequence CDR1 according to Chothia

(238) SEQ ID NO 226: Antibody 131B4-2 VH sequence CDR2 according to Chothia

(239) SEQ ID NO 227: Antibody 131B4-2 VH sequence CDR3 according to Chothia

(240) SEQ ID NO 228: Antibody 131B4-2 VK sequence CDR1 according to Chothia

(241) SEQ ID NO 229: Antibody 131B4-2 VK sequence CDR2 according to Chothia

(242) SEQ ID NO 230: Antibody 131B4-2 VK sequence CDR3 according to Chothia

(243) SEQ ID NO 231: Antibody 107C6 VH sequence CDR1 according to Kabat

(244) SEQ ID NO 232: Antibody 107C6 VH sequence CDR2 according to Kabat

(245) SEQ ID NO 233: Antibody 107C6 VH sequence CDR3 according to Kabat

(246) SEQ ID NO 234: Antibody 107C6 VK sequence CDR1 according to Kabat

(247) SEQ ID NO 235: Antibody 107C6 VK sequence CDR2 according to Kabat

(248) SEQ ID NO 236: Antibody 107C6 VK sequence CDR3 according to Kabat

(249) SEQ ID NO 237: Antibody 108F8 VH sequence CDR1 according to Kabat

(250) SEQ ID NO 238: Antibody 108F8 VH sequence CDR2 according to Kabat

(251) SEQ ID NO 239: Antibody 108F8 VH sequence CDR3 according to Kabat

(252) SEQ ID NO 240: Antibody 108F8 VK sequence CDR1 according to Kabat

(253) SEQ ID NO 241: Antibody 108F8 VK sequence CDR2 according to Kabat

(254) SEQ ID NO 242: Antibody 108F8 VK sequence CDR3 according to Kabat

(255) SEQ ID NO 243: Antibody 109A6 VH sequence CDR1 according to Kabat

(256) SEQ ID NO 244: Antibody 109A6 VH sequence CDR2 according to Kabat

(257) SEQ ID NO 245: Antibody 109A6 VH sequence CDR3 according to Kabat

(258) SEQ ID NO 246: Antibody 109A6 VK sequence CDR1 according to Kabat

(259) SEQ ID NO 247: Antibody 109A6 VK sequence CDR2 according to Kabat

(260) SEQ ID NO 248: Antibody 109A6 VK sequence CDR3 according to Kabat

(261) SEQ ID NO 249: Antibody 111A6 VH sequence CDR1 according to Kabat

(262) SEQ ID NO 250: Antibody 111A6 VH sequence CDR2 according to Kabat

(263) SEQ ID NO 251: Antibody 111A6 VH sequence CDR3 according to Kabat

(264) SEQ ID NO 252: Antibody 111A6 VK sequence 1 CDR1 according to Kabat

(265) SEQ ID NO 253: Antibody 111A6 VK sequence 1 CDR2 according to Kabat

(266) SEQ ID NO 254: Antibody 111A6 VK sequence 1 CDR3 according to Kabat

(267) SEQ ID NO 255: Antibody 111A6 VK sequence 2 CDR1 according to Kabat

(268) SEQ ID NO 256: Antibody 111A6 VK sequence 2 CDR2 according to Kabat

(269) SEQ ID NO 257: Antibody 111A6 VK sequence 2 CDR3 according to Kabat

(270) SEQ ID NO 258: Antibody 131B4 VH sequence 1 CDR1 according to Kabat

(271) SEQ ID NO 259: Antibody 131B4 VH sequence 1 CDR2 according to Kabat

(272) SEQ ID NO 260: Antibody 131B4 VH sequence 1 CDR3 according to Kabat

(273) SEQ ID NO 261: Antibody 131B4 VH sequence 2 CDR1 according to Kabat

(274) SEQ ID NO 262: Antibody 131B4 VH sequence 2 CDR2 according to Kabat

(275) SEQ ID NO 263: Antibody 131B4 VH sequence 2 CDR3 according to Kabat

(276) SEQ ID NO 264: Antibody 131B4 VH sequence 3 CDR1 according to Kabat

(277) SEQ ID NO 265: Antibody 131B4 VH sequence 3 CDR2 according to Kabat

(278) SEQ ID NO 266: Antibody 131B4 VH sequence 3 CDR3 according to Kabat

(279) SEQ ID NO 267: Antibody 131B4 VK sequence CDR1 according to Kabat

(280) SEQ ID NO 268: Antibody 131B4 VK sequence CDR2 according to Kabat

(281) SEQ ID NO 269: Antibody 131B4 VK sequence CDR3 according to Kabat

(282) SEQ ID NO 270: Antibody 131E8 VH sequence 1 CDR1 according to Kabat

(283) SEQ ID NO 271: Antibody 131E8 VH sequence 1 CDR2 according to Kabat

(284) SEQ ID NO 272: Antibody 131E8 VH sequence 1 CDR3 according to Kabat

(285) SEQ ID NO 273: Antibody 131E8 VH sequence 2 CDR1 according to Kabat

(286) SEQ ID NO 274: Antibody 131E8 VH sequence 2 CDR2 according to Kabat

(287) SEQ ID NO 275: Antibody 131E8 VH sequence 2 CDR3 according to Kabat

(288) SEQ ID NO 276: Antibody 131E8 VH sequence 3 CDR1 according to Kabat

(289) SEQ ID NO 277: Antibody 131E8 VH sequence 3 CDR2 according to Kabat

(290) SEQ ID NO 278: Antibody 131E8 VH sequence 3 CDR3 according to Kabat

(291) SEQ ID NO 279: Antibody 131E8 VK sequence CDR1 according to Kabat

(292) SEQ ID NO 280: Antibody 131E8 VK sequence CDR2 according to Kabat

(293) SEQ ID NO 281: Antibody 131E8 VK sequence CDR3 according to Kabat

(294) SEQ ID NO 282: Antibody 131H1 VH sequence CDR1 according to Kabat

(295) SEQ ID NO 283: Antibody 131H1 VH sequence CDR2 according to Kabat

(296) SEQ ID NO 284: Antibody 131H1 VH sequence CDR3 according to Kabat

(297) SEQ ID NO 285: Antibody 131H1 VK sequence 1 CDR1 according to Kabat

(298) SEQ ID NO 286: Antibody 131H1 VK sequence 1 CDR2 according to Kabat

(299) SEQ ID NO 287: Antibody 131H1 VK sequence 1 CDR3 according to Kabat

(300) SEQ ID NO 288: Antibody 131H1 VK sequence 2 CDR1 according to Kabat

(301) SEQ ID NO 289: Antibody 131H1 VK sequence 2 CDR2 according to Kabat

(302) SEQ ID NO 290: Antibody 131H1 VK sequence 2 CDR3 according to Kabat

(303) SEQ ID NO 291: Antibody 132H4 VH sequence CDR1 according to Kabat

(304) SEQ ID NO 292: Antibody 132H4 VH sequence CDR2 according to Kabat

(305) SEQ ID NO 293: Antibody 132H4 VH sequence CDR3 according to Kabat

(306) SEQ ID NO 294: Antibody 132H4 VK sequence CDR1 according to Kabat

(307) SEQ ID NO 295: Antibody 132H4 VK sequence CDR2 according to Kabat

(308) SEQ ID NO 296: Antibody 132H4 VK sequence CDR3 according to Kabat

(309) SEQ ID NO 297: Antibody 133A6 VH sequence CDR1 according to Kabat

(310) SEQ ID NO 298: Antibody 133A6 VH sequence CDR2 according to Kabat

(311) SEQ ID NO 299: Antibody 133A6 VH sequence CDR3 according to Kabat

(312) SEQ ID NO 300: Antibody 133A6 VK sequence CDR1 according to Kabat

(313) SEQ ID NO 301: Antibody 133A6 VK sequence CDR2 according to Kabat

(314) SEQ ID NO 302: Antibody 133A6 VK sequence CDR3 according to Kabat

(315) SEQ ID NO 303: Antibody 131B4-2 VH sequence CDR1 according to Kabat

(316) SEQ ID NO 304: Antibody 131B4-2 VH sequence CDR2 according to Kabat

(317) SEQ ID NO 305: Antibody 131B4-2 VH sequence CDR3 according to Kabat

(318) SEQ ID NO 306: Antibody 131B4-2 VK sequence CDR1 according to Kabat

(319) SEQ ID NO 307: Antibody 131B4-2 VK sequence CDR2 according to Kabat

(320) SEQ ID NO 308: Antibody 131B4-2 VK sequence CDR3 according to Kabat

(321) SEQ ID NO 309: CDR section of Antibody 107C6 VH sequence CDR1

(322) SEQ ID NO 310: CDR section of Antibody 107C6 VH sequence CDR2

(323) SEQ ID NO 311: CDR section of Antibody 107C6 VH sequence CDR3

(324) SEQ ID NO 312: CDR section of Antibody 107C6 VK sequence CDR1

(325) SEQ ID NO 313: CDR section of Antibody 107C6 VK sequence CDR2

(326) SEQ ID NO 314: CDR section of Antibody 107C6 VK sequence CDR3

(327) SEQ ID NO 315: CDR section of Antibody 108F8 VH sequence CDR1

(328) SEQ ID NO 316: CDR section of Antibody 108F8 VH sequence CDR2

(329) SEQ ID NO 317: CDR section of Antibody 108F8 VH sequence CDR3

(330) SEQ ID NO 318: CDR section of Antibody 108F8 VK sequence CDR1

(331) SEQ ID NO 319: CDR section of Antibody 108F8 VK sequence CDR2

(332) SEQ ID NO 320: CDR section of Antibody 108F8 VK sequence CDR3

(333) SEQ ID NO 321: CDR section of Antibody 109A6 VH sequence CDR1

(334) SEQ ID NO 322: CDR section of Antibody 109A6 VH sequence CDR2

(335) SEQ ID NO 323: CDR section of Antibody 109A6 VH sequence CDR3

(336) SEQ ID NO 324: CDR section of Antibody 109A6 VK sequence CDR1

(337) SEQ ID NO 325: CDR section of Antibody 109A6 VK sequence CDR2

(338) SEQ ID NO 326: CDR section of Antibody 109A6 VK sequence CDR3

(339) SEQ ID NO 327: CDR section of Antibody 111A6 VH sequence CDR1

(340) SEQ ID NO 328: CDR section of Antibody 111A6 VH sequence CDR2

(341) SEQ ID NO 329: CDR section of Antibody 111A6 VH sequence CDR3

(342) SEQ ID NO 330: CDR section of Antibody 111A6 VK sequence 1 CDR1

(343) SEQ ID NO 331: CDR section of Antibody 111A6 VK sequence 1 CDR2

(344) SEQ ID NO 332: CDR section of Antibody 111A6 VK sequence 1 CDR3

(345) SEQ ID NO 333: CDR section of Antibody 111A6 VK sequence 2 CDR1

(346) SEQ ID NO 334: CDR section of Antibody 111A6 VK sequence 2 CDR2

(347) SEQ ID NO 335: CDR section of Antibody 111A6 VK sequence 2 CDR3

(348) SEQ ID NO 336: CDR section of Antibody 131B4 VH sequence 1 CDR1

(349) SEQ ID NO 337: CDR section of Antibody 131B4 VH sequence 1 CDR2

(350) SEQ ID NO 338: CDR section of Antibody 131B4 VH sequence 1 CDR3

(351) SEQ ID NO 339: CDR section of Antibody 131B4 VH sequence 2 CDR1

(352) SEQ ID NO 340: CDR section of Antibody 131B4 VH sequence 2 CDR2

(353) SEQ ID NO 341: CDR section of Antibody 131B4 VH sequence 2 CDR3

(354) SEQ ID NO 342: CDR section of Antibody 131B4 VH sequence 3 CDR1

(355) SEQ ID NO 343: CDR section of Antibody 131B4 VH sequence 3 CDR2

(356) SEQ ID NO 344: CDR section of Antibody 131B4 VH sequence 3 CDR3

(357) SEQ ID NO 345: CDR section of Antibody 131B4 VK sequence CDR1

(358) SEQ ID NO 346: CDR section of Antibody 131B4 VK sequence CDR2

(359) SEQ ID NO 347: CDR section of Antibody 131B4 VK sequence CDR3

(360) SEQ ID NO 348: CDR section of Antibody 131E8 VH sequence 1 CDR1

(361) SEQ ID NO 349: CDR section of Antibody 131E8 VH sequence 1 CDR2

(362) SEQ ID NO 350: CDR section of Antibody 131E8 VH sequence 1 CDR3

(363) SEQ ID NO 351: CDR section of Antibody 131E8 VH sequence 2 CDR1

(364) SEQ ID NO 352: CDR section of Antibody 131E8 VH sequence 2 CDR2

(365) SEQ ID NO 353: CDR section of Antibody 131E8 VH sequence 2 CDR3

(366) SEQ ID NO 354: CDR section of Antibody 131E8 VH sequence 3 CDR1

(367) SEQ ID NO 355: CDR section of Antibody 131E8 VH sequence 3 CDR2

(368) SEQ ID NO 356: CDR section of Antibody 131E8 VH sequence 3 CDR3

(369) SEQ ID NO 357: CDR section of Antibody 131E8 VK sequence CDR1

(370) SEQ ID NO 358: CDR section of Antibody 131E8 VK sequence CDR2

(371) SEQ ID NO 359: CDR section of Antibody 131E8 VK sequence CDR3

(372) SEQ ID NO 360: CDR section of Antibody 131H1 VH sequence CDR1

(373) SEQ ID NO 361: CDR section of Antibody 131H1 VH sequence CDR2

(374) SEQ ID NO 362: CDR section of Antibody 131H1 VH sequence CDR3

(375) SEQ ID NO 363: CDR section of Antibody 131H1 VK sequence 1 CDR1

(376) SEQ ID NO 364: CDR section of Antibody 131H1 VK sequence 1 CDR2

(377) SEQ ID NO 365: CDR section of Antibody 131H1 VK sequence 1 CDR3

(378) SEQ ID NO 366: CDR section of Antibody 131H1 VK sequence 2 CDR1

(379) SEQ ID NO 367: CDR section of Antibody 131H1 VK sequence 2 CDR2

(380) SEQ ID NO 368: CDR section of Antibody 131H1 VK sequence 2 CDR3

(381) SEQ ID NO 369: CDR section of Antibody 132H4 VH sequence CDR1

(382) SEQ ID NO 370: CDR section of Antibody 132H4 VH sequence CDR2

(383) SEQ ID NO 371: CDR section of Antibody 132H4 VH sequence CDR3

(384) SEQ ID NO 372: CDR section of Antibody 132H4 VK sequence CDR1

(385) SEQ ID NO 373: CDR section of Antibody 132H4 VK sequence CDR2

(386) SEQ ID NO 374: CDR section of Antibody 132H4 VK sequence CDR3

(387) SEQ ID NO 375: CDR section of Antibody 133A6 VH sequence CDR1

(388) SEQ ID NO 376: CDR section of Antibody 133A6 VH sequence CDR2

(389) SEQ ID NO 377: CDR section of Antibody 133A6 VH sequence CDR3

(390) SEQ ID NO 378: CDR section of Antibody 133A6 VK sequence CDR1

(391) SEQ ID NO 379: CDR section of Antibody 133A6 VK sequence CDR2

(392) SEQ ID NO 380: CDR section of Antibody 133A6 VK sequence CDR3

(393) SEQ ID NO 381: CDR section of Antibody 131B4-2 VH sequence CDR1

(394) SEQ ID NO 382: CDR section of Antibody 131B4-2 VH sequence CDR2

(395) SEQ ID NO 383: CDR section of Antibody 131B4-2 VH sequence CDR3

(396) SEQ ID NO 384: CDR section of Antibody 131B4-2 VK sequence CDR1

(397) SEQ ID NO 385: CDR section of Antibody 131B4-2 VK sequence CDR2

(398) SEQ ID NO 386: CDR section of Antibody 131B4-2 VK sequence CDR3

(399) SEQ ID NO 387: Antibody 131B4-2 VH variable domain sequence

(400) SEQ ID NO 388: Amino Acid Sequence of IL-18BP isoform b

(401) SEQ ID NO 389: Amino Acid Sequence of IL-18BP isoform c

(402) SEQ ID NO 390: Amino Acid Sequence of IL-18BP isoform d

EXAMPLES

(403) A. Detection of Free IL-18 Versus Complex IL-18/IL-18BP

(404) 1. Common Detection of IL-18 in Patients

(405) Human IL-18 quantification in patients is performed with ELISA assays detecting total IL-18 (both free form and IL-18BP complex). The ELISA comprises commercially available antibodies (see Table 1 below). Most common ELISA assays are performed with the pair of anti-IL-18 antibodies developed by Taniguchi et al 1997 and sold by different suppliers, namely monoclonal mouse antibody 125-2H as primary/capture antibody and monoclonal rat 159-12B as secondary/developing antibody.

(406) TABLE-US-00001 TABLE 1 Scientific publications reporting IL-18 quantifications in human patients Antibodies and commercial References Assay, disease source Wong CK et al IL-18 and IL-12 levels 1. Human IL-18 ELISA kit 2000 in plasma, Systemic from MBL, #7620 Lupus Erythematosus 2. Human IL-12 ELISA kit from R&D Systems, #DP400 Park MC et al IL-18 level in serum, Human IL-18 ELISA kit from 2004 Systemic Lupus R&D Systems same as Erythematosus MBL kit #7620 Novick D et al IL-18 and IL-18BP in 1. Two human IL-18 2001 serum, Sepsis antibodies from R&D systems (mouse monoclonal biotinylated as capture # N/A and rabbit polyclonal ruthenylated as detection # N/A) 2. Two IL-18BP antibodies developed by Interpharm and Serono that are not commercially available, clone MAb No. 582.10 as capture antibody (see above, paragraph 2.2. IL- 18BP detection in human serum and urine) and rabbit polyclonal antibody for detection Novick D et al IL-18 and IL18BP levels Same as Novick et al 2001, 2010 in serum, Systemic see previous row Lupus Erythematosus Chen DY et al IL-18 levels in serum, Human IL-18 ELISA kit from 2004 Adult Still's Disease Bender MedSystems (now eBioscience) comprising 2 human IL-18 antibodies called BMS267/2MST: 1. Monoclonal capture antibody # N/A 2. Monoclonal detection antibody labeled with biotin # N/A and reaction revealed with streptavidin-HRP

(407) 2. Estimations of Free IL-18 Levels

(408) To date, there are no reports of measured levels of free IL-18. Estimations of free IL-18 are made by extrapolation using the calculation described by Novick et al 2004 (see below). The data compares levels of IL-18 and IL-18BP in human. In these studies, researchers used the pair of commercial monoclonal anti-IL-18 antibodies 125-2H and 159-12B, where antibody 125-2H is used for capture and is known to bind the IL-18/IL-18BP complex (Argiradi et al 2009). To calculate free IL-18 in patient sera, they applied the Law of Mass Action assuming that the binding of IL-18 antibodies is reversible. The calculation is performed as follow:
K.sub.D=0.4 nM=([IL-18]×[IL-18BP])/[IL-18-IL18BP]
or [IL-18] in nM=(0.4×[IL-18-IL18BP])/[IL-18BP] Where: IL-18-IL-18BP is a complex Dissociation constant as calculated by Kim et al 2000, K.sub.D=0.4 nM Stoichiometry 1:1 in the complex IL-18-IL-18BP Concentration of IL-18 is determined by electro-chemiluminescence Concentration of IL-18BP is determined by ELISA

(409) It is important to note that the authors find large variations of free IL-18 versus the total IL-18 between patients that do not reflect the ratio of IL-18 versus IL-18BP. Interestingly, this IL18/IL-18BP ratio is not reported in the cited publications. Furthermore, anti-IL18 antibodies are not able to distinguish between free IL-18 and the complex form IL-18/IL-18BP. Finally, as described by Novick et al 2001, the anti-IL-18BP antibodies do not detect IL-18BP free form but total IL-18BP since they were reported not to block the interaction between IL-18BP and IL-18, respectively monoclonal antibodies 582.10 and 657.27. Consequently, the calculation of free IL-18 using the concentration of IL-18BP lacks accuracy. Even though encouraging, the data variation indicates that free IL-18 detection could be improved with a more appropriate assay combining antibodies specifically targeting the region of IL-18 that binds to IL-18BP.

(410) 3. Confirmation that Commonly Used Commercially Antibodies do not Detect Free IL-18

(411) Eleven commercially available anti-IL-18 monoclonal antibodies were tested for their ability to prevent any IL-18 interaction with IL-18BP. The below data demonstrates that this is not the case and that none of the antibodies tested bind to the site of interaction between IL-18 and IL-18BP. Consequently, the detection of free IL-18 in human samples requires specific design and approaches targeting for example the IL-18 binding site/epitope to IL-18BP.

(412) The commonly used 125-2H and 159-12B antibodies were tested for both as capture and developing antibodies (see FIG. 2). The data indicates that both antibodies do not recognize the IL-18 epitope for IL-18BP and consequently provide only a quantification of total IL-18 (both forms free and complex to IL-18BP).

(413) In parallel to antibodies 125-2H and 159-12G, nine other commercial monoclonal antibodies were tested for their potential to detect free IL-18 in the same conditions as above. As described above, such antibody will be valuable to detect free IL-18 in biological samples. The list of tested commercial antibodies is given in the Table 2 below.

(414) TABLE-US-00002 TABLE 2 Tested monoclonal anti-IL-18 antibodies Company Antibody name MBL International D043-3, clone 25-2G D-045-6 159-12B biotin Santa Cruz Biotechnologies sc-13602 (1.51E3E1) sc-133127 (E-8) Abnova MAB 1308, clone mxsghk-18 MAB8223, clone SB116c1 MAB8224, cone SB116b1 MAB9935, clone 2 Millipore 04-1503 Anti-Interleukin 18 (clone CPTC-IL18-1) Lifespan LS-C137620 (clone 50008-2)

(415) The collected data indicates that none of the commercially available antibodies was able to distinguish the free IL-18 from its complex with IL-18BP.

(416) 4. ELISA Set Up to Detect Free IL-18

(417) 4.1 Capture of Free IL-18 with IL-18BP

(418) Microplate wells are coated with an appropriate volume phosphate buffer saline solution containing recombinant human IL-18BP (r-hIL-18BP) as shown in SEQ ID NO 7, and less than 30% of N-terminal and/or C-terminal deletion variants of IL-18BP. A description of the r-hIL-18BP is provided in section 6.6.1 below. A description of the purification protocol is provided in section 6.6.2. Plates are incubated for a period of time at 4° C. and then stabilized with a blocking buffer containing bovine serum albumin or other appropriate blocking agents. Once the reaction is finished, microplates are sealed and stored at 4° C. until used for detection of free IL-18. Microplates can also be dried in a stabilizing solution allowing storage at room temperature and then be reconstituted by hydration when needed for assay.

(419) As an example, for a final reaction volume of 100 μl, dispense first 80 μl of biotin/antibody conjugate. Samples or biological fluids containing free IL-18 are tested with the IL-18BP coated microplates. After that, 20 μl sample volume containing biological fluid or standard is dispensed per microplate well. Non-diluted or diluted biological fluid can be but is not restricted to serum, urine, tear, saliva, bile, sweat, exhalation or expiration, sputum, bronchoalveolar fluid, sebum, cellular, gland, mucosa or tissue secretion, biopsy, homogenized tissue. The free IL-18 standard concentrations range between 4.2 pg/ml to 3000 pg/ml. Standard and concentrations were prepared from commercially available recombinant human IL-18. The plates are sealed and then incubated under gentle shaking for free IL-18 capture. A suitable period of time is allowed for the reaction ranging from minutes to hours at room temperature, 37° C. or other temperatures that do not affect the stability of the samples and reagents. The microplate wells are then washed extensively with the appropriate buffer and then, 100 μl buffer developing mixture is added to each well. The developing mixture contains a streptavidin-conjugated enzyme such as peroxidase or alkaline phosphatase. The microplate wells are sealed and the reaction is allowed for a suitable period of time ranging from minutes to hours at room temperature, 37° C. or other temperatures that do not affect the stability of the samples and reagents. The resulting reactions are then monitored with a microplate reader at an appropriate nanometer wavelength for absorbance or fluorescence of the produced reagent.

(420) 4.2 Capture of Free IL-18 with Anti-IL-18 Antibody

(421) Microplate wells are coated with an appropriate volume phosphate buffer saline solution containing an antibody of the invention. Plates are incubated for a period of time at 4° C. and then stabilized with a blocking buffer containing bovine serum albumin or other appropriate blocking agents. Once the reaction is finished, microplates are sealed and stored at 4° C. until used for detection of free IL-18. Microplates can also be dried in a stabilizing solution allowing storage at room temperature and then be reconstituted by hydration when needed for assay.

(422) As an example, for a final reaction volume of 100 μl, dispense first 80 μl of biotin/antibody conjugate. Samples or biological fluids containing free IL-18 are tested with the IL-18BP coated microplates. After that, 20 μl sample volume containing biological fluid or standard is dispensed per microplate well. Non-diluted or diluted biological fluid can be but is not restricted to serum, urine, tear, saliva, bile, sweat, exhalation or expiration, sputum, bronchoalveolar fluid, sebum, cellular, gland, mucosa or tissue secretion, biopsy, homogenized tissue. The free IL-18 standard concentrations range between 4.2 pg/ml to 3000 pg/ml. Standard and concentrations were prepared from commercially available recombinant human IL-18. The plates are sealed and then incubated under gentle shaking for free IL-18 capture. A suitable period of time is allowed for the reaction ranging from minutes to hours at room temperature, 37° C. or other temperatures that do not affect the stability of the samples and reagents. The microplate wells are then washed extensively with the appropriate buffer and then, 100 μl buffer developing mixture is added to each well. The developing mixture contains a streptavidin-conjugated enzyme such as peroxidase or alkaline phosphatase. The microplate wells are sealed and the reaction is allowed for a suitable period of time ranging from minutes to hours at room temperature, 37° C. or other temperatures that do not affect the stability of the samples and reagents. The resulting reactions are then monitored with a microplate reader at an appropriate nanometer wavelength for absorbance or fluorescence.

(423) 4.3 Titration of Free IL-18 as a Function of IL-18BP Level

(424) A constant quantity of recombinant IL-18 was titrated as a function of different and well defined quantities of IL-18BP in order to understand when free IL-18 is not any more detectable. A PBS solution of 400 pg/mL IL-18 supplemented by 5% BSA was spiked with defined quantities of IL-18BP ranging from 0 to 10′000 pg/mL. The molar ratios were calculated according to the respective molecular weight of IL-18 and IL-18BP. The free IL-18 detection was performed with ELISA using IL-18BP for IL-18 as described above. The collected data presented in FIG. 1 indicates that 400 pg/mL IL-18 detection are near background detection level when IL-18BP concentration is equal or higher to 6000 pg/mL representing a molar ratio IL-18BP/IL-18 of ˜15 fold higher IL-18BP. In contrast, when molar ratio is lower than 15, free IL-18 is easily detectable.

(425) 4.4 Revised Calculation of Dissociation Constant (K.sub.D) Between Human IL-18 and IL-18BP

(426) 4.4.1 K.sub.D Calculation by Titration

(427) A K.sub.D of 400 pM is reported in the literature based on BIAcore measurements (Kim et al 2000.sup.8) However, due to the above results, the K.sub.D was revisited with the above ELISA set up. Titration of 10 pM IL-18 was performed with increasing concentrations of IL-18BP (60 pM-3 nM) in either a) healthy volunteer sera depleted in endogenous IL-18BP or b) PBS supplemented by 5% BSA. The free IL-18 ELISA in addition to commercially available assays for total IL-18 and total IL-18BP allows the determination of K.sub.D in solution which should reflect better the affinity of IL-18 to its binding protein in body fluids than data from solid-phase BIAcore method. Example of results are exposed in Table 3.

(428) TABLE-US-00003 TABLE 3 Titration of IL-18 in serum or 5% BSA solution containing 1.87 nM IL-18BP IL-18 Titration IL-18 IL-18 spiked spiked Final into into 5% Standard curve IL18 serum BSA pg/mL OD450 spiked OD450 OD450 IL-18 nm ng/mL nm nm nM IL-18 nM IL-18BP 2000 2.894 24 0.474 1.151 1.3953 1.87 666.7 2.292 20 0.342 0.897 1.1628 1.87 222.2 0.875 16 0.286 0.735 0.9302 1.87 74.1 0.303 12 0.200 0.511 0.6977 1.87 24.7 0.114 8 0.157 0.348 0.4651 1.87 8.2 0.061 4 0.091 0.188 0.2326 1.87 2.7 0.042 2 0.065 0.155 0.1163 1.87 0 0.039 0 0.040 0.037 0 1.87

(429) K.sub.D was calculated based on the following formula:
K.sub.D=[free IL-18]×[free IL-18BP]/[IL-18/IL-18BP complex]
[free IL-18BP]=[total IL-18BP]−[free IL-18]
[IL-18/IL-18BP complex]=[total IL-18]−[free IL-18]

(430) Result:

(431) K.sub.D=50 pM (Serum diluent); 35 pM (5% BSA diluent)

(432) The titration result indicates a K.sub.D of respectively 50 pM in serum diluent and 35 pM in PBS supplemented by 5% BSA. In contrast to the previous estimations of the K.sub.D between human IL-18BP and IL-18, the newly calculated K.sub.D indicates that previous estimations of free IL-18 based on the K.sub.D of 400 pM reported by Kim et al 2000 are not accurate.

(433) 4.4.2 K.sub.D Estimation by BIAcore

(434) Following the above K.sub.D results obtain by titration, we tested the binding affinity of IL-18BP to IL-18 with a simpler BIAcore setup consisting of binding IL-18BP to the BIAcore chip and then testing its affinity to IL-18. The method setup is the contrary of Kim et al 2000.sup.8, who bound IL-18 to the BIAcore chip with a monoclonal antibody and then tested the affinity of the complex antibody-IL-18 to IL-18BP. Importantly, the new BIAcore setup collected data that are aligned completely to the above titration findings, i.e. a K.sub.D ranging between 20 and 30 pM. The data is resented in Table 4 below.

(435) TABLE-US-00004 TABLE 4 New BIAcore estimation of human IL-18BP affinity to human IL-18 K.sub.a (10.sup.+5/Ms) K.sub.d (10.sup.−6 1/s) K.sub.D (10.sup.−11 M) 5.3 ± 1.2 13.3 ± 2.7 25.9 ± 4.8

(436) 4.5 Titration of Spiked IL-18 in Serum or 5% BSA Solution Containing IL-18P

(437) Human serum contains significant levels of endogenous as well as complexed IL-18 to IL-18BP, respectively at ng/mL and pg/mL levels. Both are detectable with commercially available antibodies. However, no commercially available assays are available to detect free IL-18. In order to verify the above ELISA setup for the detection of free IL-18, we spiked recombinant human IL-18 in human serum to find levels of detection. For this, nanograms of IL-18 were spiked in either serum containing endogenous 35 ng/mL IL-18BP or PBS solution supplemented by 5% BSA and 35 ng/mL IL-18BP. Resulting free IL-18 was monitored with the ELISA procedure described above. Results are presented in Table 5 below.

(438) TABLE-US-00005 TABLE 5 Spiked IL-18 detection in serum or 5% BSA containing 35 ng/ml IL-18BP IL-18 Titration IL-18 spiked IL-18 spiked Standard curve into serum into 5% BSA pg/mL OD450 Final IL18 spiked OD450 OD450 IL-18 nm ng/mL nm nm 2000 3.171 100 3.5 3.5 666.7 1.388 80 3.5 3.5 222.2 0.477 70 2.37 3.5 74.1 0.183 60 0.99 3.37 24.7 0.085 50 0.68 2.05 8.2 0.050 40 0.46 1.17 2.7 0.043 30 0.298 0.75 0 0.043 20 0.185 0.44 10 0.11 0.16 5 0.06 0.09 2 0.05 0.07 0 0.04 0.04

(439) 4.6 Detection of Free IL-18 in Serum from Patients Suffering from Adult Onset Still's Disease

(440) Following the results and in contrast to the above indications having reasonably low levels of total IL-18, we tested Adult onset Still's Disease patient samples which is known for its elevated levels of total IL-18 in serum (Kawashima et al 2001 and Chen et al 2004). As described by Kawashima et al 2001 and elsewhere, elevated total IL-18 serum levels correlate with Adult onset Still's Disease activity such as a)pyrexia, arthralgia, arthritis, cartilage damage, b) higher levels of Ferritin and c) liver enzymes (LDH). Thanks to the above ELISA set up, we report for the first time free IL-18 levels in Adult onset Still's Disease patients (see Table 6). As for the other tested indications, calculated free IL-18 levels do not correspond to the detected free IL-18 levels. The collected data indicates at least 70% of patients were positive to free IL-18.

(441) TABLE-US-00006 TABLE 6 Detection of free IL-18 in AoSD patient serum and synovial fluid Calculated Sample Total Free Free IL-18 Patient collection Biological IL-18 IL-18 pg/ml IL-18BP number date fluid pg/ml pg/ml K.sub.D = 4 × 10.sup.−10M ng/ml 1 Serum  6699 9.6 1366.5 32.6 1 Synovial   439 15.8 439 — fluid 2 Serum   713 22.5 564.3 2.0 3 Serum 106026 3.2* 59030 50.4 4 Serum 225456 24.9 157207 68.1 5 Serum 175589 23.6 139614 36.1 6 Serum  35045 2.5* 8908 45.6 7 Serum  17714 22.4 634.8 206.0 7 Synovial 133325 21.3 11162 193.6 fluid 8 Serum  25020 21.1 1277.4 153.7 9 Serum  3625 24.9 394.7 60.8 10 17 Feb. 2006 Serum  11401 7.7 6062 11.3 10 11 Jun. 2007 Serum  79942 31.6 62035 19.1 10  6 Apr. 2009 Serum  37372 18.9 22252 19.2 10  .sub. 6 Aug. 2010 Serum 185157 12.1 10566 282.9 10  6 Jun. 2012 Serum 131561 11.2 4091 341.2 11  3 Jan. 2006 Serum 150669 34.3 114012 37.2 11  4 Apr. 2007 Serum 106026 26.2 63543 45.2 11 20 Oct. 2008  Serum 225456 23.6 70633 163.0 11 21 Apr. 2010  Serum 175589 23.3 116583 59.8 12 20 Jun. 2009  Serum  3625 8.0 1633 10.5 13 10 Mar. 2010  Serum   439 4.8** 151.2 13.7 14 17 Jul. 2009   Serum 133325 19.3 21118 144.4 15 24 Jul. 2006   Serum  35045 14.3 14628 29.3 16 25 Apr. 2007  Serum  17714 8.0 4075 36.6 16 10 Jun. 2010   Serum  25020 6.4 2592 82.4 *: Level comparable to the background signal **: Level comparable to the lower limit of detection —: not detectable, level comparable to the background signal

(442) 5. Conclusions

(443) The data in both publications and the above experimental setup demonstrate that commercial monoclonal antibodies detect total IL-18 but not free IL-18. Furthermore, the most commonly used antibodies to quantify IL-18, namely 125-2H and 159-12B, are confirmed as well in detecting total IL-18.

(444) The estimation of free IL-18 using the Law of Mass Action is an interesting approach. Nevertheless, the large error bars obtained do not support its use in clinical monitoring. Furthermore, the anti-IL-18BP antibodies detect total IL-18BP and not the free form. Consequently, the calculation of free IL-18 using the concentration of IL-18BP lacks accuracy.

(445) The proposed approach to quantify free IL-18 by targeting IL-18 binding site to IL-18BP seems more appropriate and is demonstrated for the first time to be more accurate than extrapolated quantifications with the Law of Mass Action. In addition, the affinity of IL-18BP is higher than reported by Kim et al 2000 with a K.sub.D ranging near 50 pM in serum and 20-30 pM with a new BIAcore setup.

(446) 6. Administrations of IL-18BP in Patients with Adult-Onset Still's Disease (AoSD)

(447) 6.1 Objectives

(448) Primary:

(449) To assess the safe use of r-hIL-18BP in AoSD patients

(450) Secondary:

(451) To assess clinical efficacy and laboratory/biological evidence of efficacy

(452) 6.2 Number of Patients

(453) 30 patients

(454) 6.3 Inclusion Criteria Patients aged 18 years and older, diagnosed as AoSD based on the presence of the Yamaguchi criteria (see appendix 2) with active disease, irrespective of the continuation of the permitted treatment mentioned below. Patients with active disease will be considered if they exhibit at least two of the Yamaguchi's major criteria (see appendix 2) at the screening visit plus at least either fever or elevation of markers of inflammation (CRP ≥10 mg/L and/or Erythrocyte Sedimentation Rate ESR ≥28 mm/h). Patients that have been exposed to NSAIDS, Prednisone (at least 5 mg/day) for ≥1 month) and/or synthetic sDMARDs (methotrexate at a dose of at least 10 mg/week) for ≥3 months without response to treatment or with incomplete response to treatment Women of childbearing potential with negative pregnancy test at screening, V3, V4, V5 and V6 and that agree to follow highly effective birth control recommendations during the study and until 1 month after the end of the treatment. Birth control methods that are considered as highly effective are either: combined (estrogen and progestogen containing) hormonal contraception associated with inhibition of ovulation, progestogen-only hormonal contraception associated with inhibition of ovulation, intrauterine device (IUD), intrauterine hormonereleasing system (IUS), bilateral tubal occlusion, vasectomized partner or sexual abstinence. In each case of delayed menstrual period (over one month between menstruations, confirmation of absence of pregnancy is strongly recommended. This recommendation also applies to women of child bearing potential with infrequent or irregular menstrual cycles. As regards the duration of contraception after the study, taking into account the median half-life of r-hIL-18BP of almost 40h, 5 half-lives represent duration of 200 hours. In order to be on the safe side, a post-study contraception duration of 4 weeks is recommended Patients can maintain treatment with stable doses of Non-Steroidal anti-inflammatory Drugs (NSAIDs), Prednisone (stable dose of Prednisone (of at least 5 mg/day), and sDMARDs during r-hIL-18BP treatment (methotrexate at a dose of at least 10 mg/week). Specifically baseline levels of prednisone treatment can be maintained or tapered (due to patient improvement), any requirement for prednisone increase during treatment will be considered a treatment failure. Ability to understand and willingness to sign a written informed consent Previous treatments with biologicals are allowed if the following washout periods are respected: one week for anakinra, two weeks for etanercept, and 6 weeks for adalimumab, certolizumab, golimumab, tocilizumab, abatacept and 8 weeks for infliximab. Previous rituximab administration will require 6 months of washout and normal B-cell counts and previous treatment with canakinumab will require 6 months of washout.

(455) 6.4 Exclusion Criteria Patients with a first episode of AoSD with less than one month of therapy with Prednisone or sDMARDs Patients with active or chronic infections (i.e. Tuberculosis (TB), HIV, HBV & HCV) Patients suffering from inherited immunodeficiency diseases Patients with white blood cell counts below 2′500 cells/mm.sup.3 Patients with Neutrophils below 1′000 cells/mm.sup.3 Concomitantly treated with biologicals Women of childbearing potential who are unwilling to use adequate protection from pregnancy Women of childbearing potential who are unwilling to use highly effective birth control methods (see definition in Inclusion criteria above) up to 1 month after the end of her participation in the study. Inability to understand and unwilling to sign a written informed consent Active Macrophage Activating Syndrome (MAS) Any acute or chronic life-threatening disease Such as cancer, and irreversible organ failures of heart, liver, lung and kidney (creatinine not higher than 1.5×upper limit of normal). Patients having received adalimumab, certolizumab, golimumab, tocilizumab and abatacept within 6 weeks, infliximab within 8 weeks, canakinumab within 6 months, etanercept within 2 weeks, or anakinra 1 week prior to the start of r-hIL-18BP will not be enrolled into the study. Patients that have received rituximab within 6 months and/or have persistent low B-cell counts will not be eligible for enrolment. Subject who cannot be expected to comply with the study procedures Currently participating or having participated in another clinical trial during the last 4 weeks prior to the beginning of this study. Patients with a history of severe hypersensitivity reactions

(456) 6.5 Study Duration

(457) Twelve (12) week treatment will be followed by a 4-week follow-up period for safety assessments. Data management, statistical and study report will take 4 more months

(458) 6.6 Study Drug

(459) 6.6.1 Description

(460) r-hIL-18BP is a soluble glycoprotein of 164 amino acids produced from a Chinese Hamster Ovary cell line. The polypeptide chain contains 6 cysteine residues located at positions 21, 34, 56, 59, 101 and 120, that are theoretically predicted to form three disulphide bond pairs. The molecule contains four potential N-glycosylation sites at asparagine 49, 64, 73 and 117. The average molecular weight of the full-length polypeptide moiety of r-hIL-18BP calculated on the basis of the amino acid composition is around 17.6 kD. The relative molecular mass of the whole molecule is approximately 50 kDa (including glycans).

(461) 6.6.2 Purification Protocol

(462) The purification process starts with the removal of cells and cell debris from the harvested cell culture supernatant by centrifugation, diafiltration and transfer into a mixing tank. The harvest containing non-homogenous IL-18BP is concentrated and diafiltered against a basic borate buffer. After that, IL-18BP is captured on TMAE Hi-Cap anion-exchange resin to remove salts and cell culture nutrients. The IL-18BP is eluted with a basic borate buffer supplemented by NaCl.

(463) Five additional chromatographic steps are performed to homogenize IL-18BP, including two tangential flow filtration steps and a virus removal filtration step as follow: The protein preparation is processed through Immobilized Metal Affinity Chromatography on Chelating Sepharose Fast Flow resin, activated with copper, to remove host cell proteins. The protein is eluted with ammonium acetate. The Immobilized Metal Affinity Chromatography eluate is loaded onto the hydrophobic charge induction chromatography column on MEP HyperCel to remove further host cell proteins. The product is eluted with an alkaline phosphate buffer containing propylene glycol. The eluate containing IL-18BP is then concentrated by diafiltration. The retentate of the diafiltration is diluted and adjusted to acidic pH with 2-(N-morpholino)ethanesulfonic acid (MES) buffer. After that, the protein is separated by ion-exchange chromatography by loading onto a CM Sepharose Fast Flow column in flow-through mode to remove remaining host cell proteins, which are retained on the column. The unbound fraction is adjusted to basic pH with sodium tetraborate. The fraction from the ion exchange chromatography step is then separated by hydrophobic interaction chromatography column on Phenyl Sepharose Fast Flow for further polishing. The column is pre-equilibrated with borate buffer containing a high molarity of ammonium sulfate, and the product is eluted from the column by lowering the concentration of ammonium sulfate. Subsequently, a nanofiltration step follows to ensure a proper removal of virus particles. The permeate is processed by Reverse Phase chromatography on Source 30 RPC resin as the final polishing step. Elution of the product is achieved through an acetonitrile gradient supplanted by TFA 0.1% The eluate containing homogenized IL-18BP is concentrated by an ultrafiltration/diafiltration step and finally filtered through 0.22 μm filter for storage.

(464) All purification steps are performed at room temperature.

(465) The final composition obtained is substantially free of N-terminal and/or C-terminal deletion variants of IL-18BP and contains between 2% and 8%, but less than 10% of said variants.

(466) 6.6.3 Composition

(467) The drug product formulation (recombinant human interleukin 18 Binding protein (rhIL-18BP) will have a strength of 80 mg, and is prepared in a sterilized solution for injection containing sodium chloride, 7 mg per vial, sodium dihydrogen phosphate monohydrate approx. 1.0 mg per vial, disodium phosphate dehydrate approx. 2.4 mg per vial, sodium hydroxide and 0-phosphoric acid 85% to adjust to pH 7.0, water for injection up to 1 ml. (See appendix 3).

(468) Glass vials containing 1 ml of the injection volume and 80 mg of the recombinant molecule will be the administration unit.

(469) 6.7 Dose/Route Regime

(470) 80 mg, 160 mg and 320 mg/Subcutaneous/Three times a week (TIW).

(471) Patients will receive the treatments three times a week (TIW). The 80 mg cohort will receive 1 ml of the study product. Patients of the 160 mg dose cohort will receive 2 vials and those of the 320 mg cohort, 4 vials. A volume of 1 ml will be withdrawn from each vial using a separate 2 ml sterile, single use syringe for each vial. Before injection, vials will be brought to room temperature (18-25° C.) by removing them from the refrigerator 30 min prior to administration.

(472) 6.7.1 Drug Product Selected Dose:

(473) No validated animal model has been described for this condition. In the collagen-induced arthritis murine model the active dose for r-hIL-18BP administered intraperitoneally (i.p.) was determined to be 1 mg/Kg body weight. Since the bioavailability by the i.p. route is almost of 100% while the s.c. route attains 55% an adjustment of the s.c. dose by a factor of 0.55 was calculated and thus the effective s.c. dose in mice would result in 1.82 mg/kg. If the difference between human and mouse dissociation constant (Kd) is taken into account (0.16 nM v. 0.40 nM) a further correction factor of 2.67 fold should be introduced. Altogether the expected pharmacologically active dose for a “humanized mice” treated by s.c. route with r-hIL-18BP is round 4.85 mg/kg. Translation of this dose to human using the accepted allometric murine conversion factor of 12.3 (as per the 2005 US Guidance “Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers”) reveals a human equivalent dose of 0.39 mg/Kg, or 27.3 mg for a 70 Kg individual. Although the guidance was initially written for extrapolating animal doses to healthy volunteers, its principles can be applied also to patients. From a clinical efficacy point of view it is important to underline that doses of 20 mg 3 times a week showed no efficacy in RA and Pso patients. Those doses will be likely also ineffective in AoSD patients considering that blood levels of IL-18 in AoSD are higher than those reported in the two above mentioned conditions. In conclusion, based on preclinical and previous human studies performed with r-hIL-18BP and in order to avoid providing inefficient doses to AoSD patients during the active phase of the disease, a dose-escalating study with a starting dose of 80 mg TIW and a step wise progression to 160 and 320 mg is proposed in the present clinical trial.

(474) 6.7.2 Route of Administration:

(475) The product comes as a solution to be administered by the s.c. route. Glass vials contain 1 ml with a 15% overfill with a dose of 80 mg of the active product. Patients in the 160 mg and 320 mg cohorts will receive separate s.c. injections of 1 ml each: 2 injections of 1 mL for the 160 mg dose and 4 injections of 1 mL for the 320 mg dose. A separate syringe must be used for each injection.

(476) The site of the s.c. injection should be alternated e.g. the outside of the thighs and the various quadrants of the anterior abdominal wall. The separate injections that constitute a single dosage of study drug should be administered within the same body region but not at the exact same injection site.

(477) 6.8 Methodology

(478) Patients will be allocated to three cohorts that will receive doses of 80 mg, 160 mg or 320 mg. Patients in cohort 1 (n=10) will receive injections of 80 mg RhIL-18BP TIW by the subcutaneous route. After the first 5 patients have completed three weeks of treatment a Data Safety Monitoring Board (DSMB) will perform a safety evaluation. If a cohort shows 2 or more Severe or Serious Adverse events (SAEs) the DSMB will decide on the interruption or not of the trial at this dose. The DSMB will give the authorization to pursue enrollment up to ten patients and to open the next upper dose cohort. Patients in cohort 2 (n=10) will receive injections of 160 mg rhIL-18BP TIW by the subcutaneous route. After the first 5 patients have completed three weeks of treatment the DSMB will perform a safety evaluation. If a cohort shows 2 or more SAEs the DSMB will decide on the interruption or not of the trial at this dose. The DSMB will give the authorization to pursue enrollment up to 10 patients and to open the next upper dose cohort. Patients in cohort 3 (n=10) will receive injections of 320 mg rhIL-18BP TIW by the subcutaneous route. After the first 5 patients have completed three weeks of treatment the DSMB will perform a safety evaluation. If the cohort shows 2 or more SAEs the DSMB will decide on the interruption or not of the trial at this dose. The DSMB will give the authorization to enroll 5 more patients until the completion of the treatment.

(479) Patient's visits are scheduled for week −2 (screening visit and signature of ICF), week 0 (study entry visit, beginning of treatment) weeks 1 (V2), 3 (V3), 6 (V4), 12 (V5) and 16 (V6). End of treatment occurs at 12 weeks (V5) and patients are followed for safety and tolerability for a further one month period (V6). For patients that are non-responders to a given dose after 3-week treatment and are titrated up to the next upper dose the treatment duration will be of 15-weeks followed by 4 weeks for safety evaluation.

(480) 6.9 Statistical Analysis

(481) The sample size has been defined according to the commonly accepted studies in orphan rare disease. Safety and tolerability of rhIL-18BP treatment will be assessed by the incidence and outcome of AEs, the routine clinical examination and the safety laboratory tests. The incidence of AEs will be described according to the MedDRA System Organ Class (SOC) and Preferred Term (PT) for all events, by severity and relationship to study treatment. Laboratory parameters (haematology and chemistry) will be summarised over the scheduled protocol visits in terms of actual values and changes from Baseline. In addition, shift tables will be produced comparing baseline to final non-missing values Descriptive statistics will be used to summarize demographic and baseline characteristics. Selected variables at 3 and 12 weeks will be compared to baseline values using paired t-test or Wilcoxon's signed rank test. The principal evaluation of efficacy (key secondary end-point) is at 12 weeks. For selected variables the mean±SEM and 95% confidence intervals will be calculated from all data points available for the respective time point (3 or 12 weeks). Dose efficacy will be considered attained if more than 50% of the patients show a positive response to treatment. The association between the patient response to treatment and the levels of free IL-18 levels at baseline will be studied. The pharmacokinetic study of rhIL-18BP will include the following end-points: Time to maximal concentration (tmax), the maximal concentration (Cmax), trough concentration, area under the curve during the first 24h treatment (AUC 0-24) and the elimination half life will be determined in serum. The pharmacodynamics study will take into account direct PD end-points: free and total IL-18 and as indirect PD end-points: CRP and IL-6 as a function of time and drug treatment.

(482) 7. IL-18BP Efficacy in COPD Exacerbation Mouse Model

(483) The aim of the study was to determine the effect of IL-18BP, administered at three dose levels, by the sub-cutaneous route, on Polyinosinic:polycytidylic acid-induced exacerbation of tobacco-smoke induced pulmonary inflammation, in C57BL/6J mice. High level of dexamethasone, dosed orally, was included in the study as a reference agent.

(484) 7.1 IL-18BP Composition

(485) The IL-18BP is r-hIL-18BP administered as a sterilized injectable solution. A description of the experimental drug is provided in section 6.6.1 above. A description of the purification protocol is provided in section 6.6.2 and the composition of the injectable solution in section 6.6.3.

(486) 7.2 General Methodology: Four-Day Exacerbation/Tobacco Smoke Mouse Model

(487) Mice received either vehicle (PBS) or IL-18BP. IL-18BP was given subcutaneously to 3 groups of animals respectively at 1, 3 or 10 mg/kg 2h prior to the initial tobacco smoke exposure from Day 1 to Day 4. Mice received orally either vehicle or dexamethasone (10 mg/kg) 1h prior to each twice daily exposure. Mice received by intranasal administration either the vehicle or Polyinosinic:polycytidylic acid (2 mg/kg) 2h prior to the initial air or tobacco smoke exposure on Day 4 to induce lung inflammation exacerbation. Tobacco smoke exposure was performed during the morning and afternoon as follow: Day 1 for 15 min, Day 2 for 25 min, Day 3 for 30 min and Day 4 for 30 min.

(488) Animal groups and their respective treatment regimes are summarized in Table 6.1.

(489) TABLE-US-00007 TABLE 6.1 Treatment regimes for tobacco smoke mouse model Treatment Treatment Dose Exposure s. c. /oral Code n mg/kg Challenge Frequency Air Veh/Veh A 10 —/— Veh Sub- cutaneous TS Veh/Veh B 10 —/— Veh 2h prior to initial TS Air Veh/Veh C 10 —/— p[I:C] on each 2 mg/kg day TS Veh/Veh D 10 —/— p[I:C] Oral 1h 2 mg/kg prior to TS IL-18BP/Veh E 10 1/— p[I:C] each TS 2 mg/kg exposure TS IL-18BP/Veh F 10 3/— p[I:C] on each day 2 mg/kg TS IL-18BP/Veh G 10 10/— p[I:C] p[I:C] 2 mg/kg intranasal TS Veh/Dex H 10 —/10 p[I:C] 2h prior to 2 mg/kg TS exposure on day 4 TS: Tobacco smoke; Veh: Vehicule; Dex: Dexamethazone, p[I:C]: Polyinosinic:polycytidylic acid

(490) Following the above treatments, animals were terminally anaesthetised on Day 5. After that, a blood sample was taken via the sub-clavian artery (plasma) and the animals were bronchoalveloar lavaged with 3×0.4 ml of PBS for further cellular and cytokine/mediator analysis. Bronchoalveolar lavage supernatants were stored at −80° C. for cytokine/mediator analysis. Cells recovered from the BALF were counted using the Sysmex cell counter. Finally, the collected data was statistically analyzed by Students t-test and ANOVA (Sidak's was used in the case of data passed normality test or Kruskal Wallis test if data did not pass normality test).

(491) 7.3 Confirmation of IL-18 Pathway Activation in the Four-Day Exacerbation/Tobacco Smoke Mouse Model

(492) Mouse IL-18 was tested in the BAL using a commercial ELISA in order to confirm the mouse model for IL-18 pathway activation. The collected data indicates a clear induction of IL-18 in the lung airway space (see FIG. 5). IL-18 is not detectable in the control (air only). Interestingly, IL-18 is expressed under smoke exposure but is not significantly induced under poly[I:C] alone (under the lower limit of detection). In contrast and as expected, the combination of smoke and poly[I:C] raises considerably IL-18 to much higher levels in the BAL than smoke or poly[I:C] alone.

(493) 7.4 Exacerbated Inflammation Mitigation by IL-18BP in Exacerbation/Tobacco Smoke Mouse Model

(494) 7.4.1 Inhibition of Total Cell Infiltration and Exacerbated Inflammation in the Lung Airway Space by IL-18BP

(495) Mice treated by IL-18BP had a significant mitigation of total cell infiltration in the lung following induction of exacerbated inflammation. Doses of either 3 and 10 mg/kg indicated statistically valuable efficacy compared to the positive control dexamethasone (see FIG. 5). It is important to note that dexamethasone had no sign of efficacy at 3 mg/kg doses in the mouse model (data not shown), indicating that the high dexamethasone dose of 10 mg/kg is potentially inducing apoptosis in certain cell types such as macrophages, eosinophils and lymphocytes (data not shown). Similar observation was made with Roflumilast [3-(cyclopropylmethoxy)-N-(3,5-dichloropyridin-4-yl)-4-(difluoromethoxy) benzamide] in the mouse model where no hint of cell infiltration inhibition was observed with 2.5 mg/kg dose (data not shown). FIG. 6 shows clear and statistically relevant efficacy of IL-18BP at 10 mg/kg in exacerbated inflammation inhibition in the current mouse model.

(496) 7.4.2 Inhibition of Neutrophil Infiltration in the Lung Airway Space by IL-18BP

(497) Neutrophil infiltration was inhibited by IL-18BP in tobacco smoke-exacerbated lungs. Doses of either 3 and 10 mg/kg IL-18BP indicated statistically valuable efficacy compared to the positive control dexamethasone (see FIG. 7). In the current mouse model conditions, IL-18BP 10 mg/kg dose seems to have the best statistical efficacy (see FIG. 8).

(498) 7.4.3 Inhibition of Granulocyte Colony-Stimulating Factor (G-CSF) Pathway in the Lung Airway Space by IL-18BP

(499) G-CSF is well acknowledged as key cytokine stimulating the survival, proliferation, differentiation, and function of neutrophil precursors and mature neutrophils. Consequently, mitigation of G-CSF pathway-induced by smoke-p[I:C] is an significant factor demonstrating an effect of IL-18BP on neutrophil recruitment in the mouse lung airway space. The presence of G-CSF in the BALF was monitored with a commercially available ELISA kit. FIG. 9 demonstrates that administration of IL-18BP mitigates G-CSF release in the lung airways, thereby confirming the inhibition of neutrophil infiltration. The three tested IL-18BP doses have a statistically relevant effect in the mouse model.

(500) 7.4.4 IL-18BP Safety: Effect on Weight Loss in Exacerbation/Tobacco Smoke Mouse Model

(501) IL-18BP administration appeared to be well tolerated by exacerbation/tobacco smoke mouse model. As an example, weight loss was mitigated by IL-18BP even though both Students t-test and ANOVA statistical analyses were not significant (see FIG. 10). A large majority of mice receiving 3 and 10 mg/kg IL-18BP lost respectively 6-7% weight in contrast to the control exposed to tobacco smoke and p[I:C] that lost about 9%. Hence, weight loss alleviation data indicates that IL-18BP is not providing additional stress to the animal model. It is interesting to note that mice receiving only p[I:C] did not lose weight compared to mice receiving the combination of p[I:C] and tobacco smoke [see FIG. 8, treatment 3) and 4)].

(502) 8. Generation of Anti-IL-18 Monoclonal Antibodies

(503) 8.1 Mouse Immunization and Monoclonal Antibody Screening

(504) Mice were vaccinated against human interleukin-18 using a technology allowing immunization with properly folded proteins. Prior to immunization, genetically modified mice were selected for major histocompatibility complexes supposedly sensitive to IL-18 surface area epitopes binding IL-18BP. Following immunization, B cells were isolated from spleen and hybridized following standard hybridoma technology. Hybridoma were sorted onto microplates and then tested for expression of monoclonal anti-IL-18 antibodies targeting IL-18 epitopes included in IL-18BP binding site. The screening was performed in 3 sequential and selective steps: First step. Positive antibody screening attempt was performed with IL-18 attached to Luminex beads confirming cell expressing monoclonal anti-IL-18 antibodies. Second step. Potential antibodies targeting IL-18 on IL-18BP binding site were rescreened in competition with IL-18BP, but not with IL-18BP fusions with Fc antibody domains, or other type of fusions, in order to prevent false antibody positive candidates due to steric hindrance created by the fused peptide. For this, monoclonal antibodies were bound to Luminex beads carrying IL-18. The complex was then exposed to biotinylated IL-18BP in order to identify interference to previously identified anti-IL-18 antibodies (see Table 7, Column #2). The second screening carried more than 300 positive antibody candidates (see Table 7, Column #3). The number of positive candidates was surprisingly high suggesting an excellent mouse immunization to the targeted epitope area. However, inhibitions were not sufficient due to diminished but still persistent fluorescence signals, thus indicating binding of IL-18BP to the complexed antibody IL-18. Nevertheless and importantly, such standard screening method reported elsewhere does not take into account a potential steric hindrance of the large antibody molecule (about 160 kDa) against the much smaller IL-18BP (about 18 kDa, peptide only). Third step. A third screening program was undertaken with Luminex beads linked to IL-18BP and then complexed to interleukin-18, assuring the presentation of properly folded recombinant IL-18 to positive antibody candidates. The resulting screening was considerably more selective because most of the above antibodies still bound the Luminex-IL-18 beads thereby indicating that their previous inhibitory effect to IL-18BP was due to steric hindrance. Finally, a total of 12 antibodies were finally considered as targeting IL-18 on the IL-18BP protein due to their very low fluorescence signal after binding IL-18 in the presence of IL-18BP, namely clone #107C6, 108F8, 109A6, 111A6, 129C3, 131B4, 131E8, 131H1, 132C12, 132H4, 133A6 and 134B2 (see Table 7, Column #4, selected clones representing inhibition means of more than 500 fold compared to Column #2). The positive antibodies versus a set of negatives are presented in Table 7 below.

(505) The collected data from third screening step (Table 7, Column #4) promoted further mRNA sequencing and clone dilution work to enrich positive monoclonal cells out of #107C6, 108F8, 109A6, 111A6, 129C3, 131B4, 131B4-2, 131E8, 131H1, 132C12, 132H4, 133A6 and 134B2. The respective KD values of each monoclonal antibody was calculated following titration of the antibodies with a defined IL-18 molarity and derived from the obtained EC.sub.50 values with the Law of Mass Action (see above). All of these monoclonal antibodies were confirmed to bind to IL-18 on the IL-18BP binding site.

(506) TABLE-US-00008 TABLE 7-1 Screening of monoclonal antibodies targeting IL-18 on the IL-18BP binding site Column #1 Column #2 Column #3 Monoclonal IL-18BP binding on Monoclonal antibody antibodies IL-18 previously binding on IL-18 binding on complexed to previously complexed to Clone IL-18 monoclonal antibody IL-18BP name Fluorescence intensity Examples of negative antibodies not following selection criteria 101D2 26 963 1 226 1 544   104H10 26 508 1 199 2 499   105A2 21 528 1 886 1 840   106H1 27 178 1 011 1 324   108F3 23 496 1 964 2 383   108G6 25 652 1 137 2 507   115E6 25 752 1 604 2 649   119E9 25 420 1 307 2 931   Positive antibodies following selection criteria 107C6 26 250 1 389 33 108F8 25 126 1 292 45 109A6 25 848   913 33 111A6 25 855 1 398 42 131B4 24 838 1 656 41 131E8 25 411 1 389 36 131H1 24 806 1 026 24 132C12 24 541 1 515 48 132H4 23 839 1 488 28 133A6 23 273 1 631 25 134B2 24 278 1 261 48 129C3 25 412   760 44

(507) TABLE-US-00009 TABLE 7-2 KD values Antibody # KD picomolar 107C6 <2 107C6 <2 107C6 <2 Mean <2 108F8 <2 108F8 <2 108F8 <2 Mean <2 109A6 10 109A6 10 109A6 10 Mean 10 111A6 5 111A6 5 111A6 5 Mean 5 129C3 <2 129C3 <2 129C3 <2 Mean <2 131B4 20 131B4 10 Mean 15 132H4 5 132H4 5 132H4 5 Mean 5 133A6 10 133A6 10 133A6 10 Mean 10

(508) TABLE-US-00010 TABLE 8  Antibody Chothia Kabat IMGT CDR-Section 107C6 VH CDR1 GYTFTNY (SEQ ID NO: NYGMN (SEQ ID NO: 231) GYTFTNYG (SEQ ID NO: 27) GYTFTNYGMN (SEQ  153) ID NO: 309) VH CDR2 NTYSGV (SEQ ID NO: 154) WINTYSGVPTYADDFKG INTYSGVP (SEQ ID NO: 28) WINTYSGVPTYADDFKG (SEQ ID NO: 232) (SEQ ID NO: 310) VH CDR3 EGYSTTRSMDY (SEQ ID EGYSTTRSMDY (SEQ ID AREGYSTTRSMDY (SEQ ID AREGYSTTRSMDY  NO: 155) NO: 233) NO: 29) (SEQ ID NO: 311) VK CDR1 KSSQSLLDSRTRKNYLV KSSQSLLDSRTRKNYLV QSLLDSRTRKNY (SEQ ID NO: KSSQSLLDSRTRKNYLV (SEQ ID NO: 156) (SEQ ID NO: 234) 30) (SEQ ID NO: 312) VK CDR2 WASTRGS (SEQ ID NO: WASTRGS (SEQ ID NO: WAS (SEQ ID NO: 31) WASTRGS (SEQ ID  157) 235) NO: 313) VK CDR3 KQSYNLRT (SEQ ID NO: KQSYNLRT (SEQ ID NO: KQSYNLRT (SEQ ID NO: 32) KQSYNLRT (SEQ ID  158) 236) NO: 314) 108F8 VH CDR1 GYTFTNY (SEQ ID NO: NYGMN (SEQ ID NO: 237)  GYTFTNYG (SEQ ID NO: 33) GYTFTNYGMN (SEQ 159) ID NO: 315) VH CDR2 NTYSGV (SQ ID NO: 160)  WINTYSGVPTYADDFKG INTYSGVP (SEQ ID NO: 34) WINTYSGVPTYADDFKG (SEQ ID NO: 238) (SEQ ID NO: 316) VH CDR3 EGYSTTRSMDY (SEQ ID EGYSTTRSMDY (SEQ ID AREGYSTTRSMDY (SEQ ID AREGYSTTRSMDY  NO: 161) NO: 239) NO: 35) (SEQ ID NO: 317) VK CDR1 KSSQSLLDSRTRKNYLV KSSQSLLDSRTRKNYLV QSLLDSRTRKNY (SEQ ID NO: KSSQSLLDSRTRKNYLV (SEQ ID NO: 162) (SEQ ID NO: 240) 36) (SEQ ID NO: 318) VK CDR2 WASTRGS (SEQ ID NO: WASTRGS (SEQ ID NO: WAS (SEQ ID NO: 37) WASTRGS (SEQ ID  163) 241) NO: 319) VK CDR3 KQSYNLRT (SEQ ID NO: KQSYNLRT (SEQ ID NO: KQSYNLRT (SEQ ID NO: 38) KQSYNLRT (SEQ ID  164) 242) NO: 320) 109A6 VH CDR1 GFKIKDT (SEQ ID NO: DTYIH (SEQ ID NO: 243) GFKIKDTY (SEQ ID NO: 39) GFKIKDTYIH (SEQ  165) ID NO: 321) VH CDR2 DPANGN (SEQ ID NO: RIDPANGNTIYGSKFQG IDPANGNT (SEQ ID NO: 40) RIDPANGNTIYGSKFQG 166) (SEQ ID NO: 244) (SEQ ID NO: 322) VH CDR3 YVWFAY (SEQ ID NO: YVWFAY (SEQ ID NO: AGYVWFAY (SEQ ID NO: 41) AGYVWFAY (SEQ ID 167) 245) NO: 323) VK CDR1 RSSQRLVHSNGNTYLH RSSQRLVHSNGNTYLH QRLVHSNGNTY (SEQ ID NO: RSSQRLVHSNGNTYLH (SEQ ID NO: 168) (SEQ ID NO: 246) 42) (SEQ ID NO: 324) VK CDR2 TVSNRFS (SEQ ID NO: TVSNRFS (SEQ ID NO: TVS (SEQ ID NO: 43) TVSNRFS (SEQ ID  169) 247) NO: 325) VK CDR3 SQSTLVPWT (SEQ ID NO: SQSTLVPWT (SEQ ID SQSTLVPWT (SEQ ID NO: 44) SQSTLVPWT (SEQ ID 170) NO: 248) NO: 326) 111A6 VH CDR1 GFKIKDT (SEQ ID NO: DTYIH (SEQ ID NO: 249) GFKIKDTY (SEQ ID NO: 45) GFKIKDTYIH (SEQ  171) ID NO: 327) VH CDR2 DPANGN (SEQ ID NO: RIDPANGNTIYGSKFQG IDPANGNT (SEQ ID NO: 46) RIDPANGNTIYGSKFQG 172) (SEQ ID NO: 250) (SEQ ID NO: 328) VH CDR3 YVWFAY (SEQ ID NO: YVWFAY (SEQ ID NO: AGYVWFAY (SEQ ID NO: 47) AGYVWFAY (SEQ  173) 251) ID NO: 329) VK1 CDR1 RARSSVSSSYLH (SEQ ID RARSSVSSSYLH (SEQ SSVSSSY (SEQ ID NO: 48) RARSSVSSSYLH  NO: 174) ID NO: 252) (SEQ ID NO: 330) VK1 CDR2 STSNLAS (SEQ ID NO: STSNLAS (SEQ ID NO: STS (SEQ ID NO: 49) STSNLAS (SEQ ID  175) 253) NO: 331) VK1 CDR3 QQYSGYPLT (SEQ ID NO: QQYSGYPLT (SEQ ID QQYSGYPLT (SEQ ID NO: 50) QQYSGYPLT (SEQ  176) NO: 254) ID NO: 332) VK2 CDR1 RSSQRLVHSNGNTYLH RSSQRLVHSNGNTYLH QRLVHSNGNTY (SEQ ID NO: RSSQRLVHSNGNTYLH (SEQ ID NO: 177) (SEQ ID NO: 255) 51) (SEQ ID NO: 333) VK2 CDR2 TVSNRFS (SEQ ID NO: TVSNRFS (SEQ ID NO: TVS (SEQ ID NO: 52) TVSNRFS (SEQ ID  178) 256) NO: 334) VK2 CDR3 SQSTLVPWT (SEQ ID NO: SQSTLVPWT (SEQ ID SQSTLVPWT (SEQ ID NO: 53)  SQSTLVPWT (SEQ  179) NO: 257) ID NO: 335) 131B4 VH1 CDR1 GFKIKDT (SEQ ID NO: DTYIH (SEQ ID NO: 258) GFKIKDTY (SEQ ID NO: 54) GFKIKDTYIH  180) (SEQ ID NO: 336) VH1 CDR2 DPANGN (SEQ ID NO: RIDPANGNTIYGSKFQG IDPANGNT (SEQ ID NO: 55) RIDPANGNTIYGSKFQG 181) (SEQ ID NO: 259) (SEQ ID NO: 337) VH1 CDR3 YVWFAY (SEQ ID NO: YVWFAY (SEQ ID NO: AGYVWFAY (SEQ ID NO: 56) AGYVWFAY (SEQ  182) 260) ID NO: 338) VH2 CDR1 GFSLTSY (SEQ ID NO: SYGVH (SEQ ID NO: 261) GFSLTSYG (SEQ ID NO: 104) GFSLTSYGVH (SEQ  183) ID NO: 339) VH2 CDR2 WRGGS (SEQ ID NO: 184) VIWRGGSTDYNAAFMS IWRGGST (SEQ ID NO: 105) VIWRGGSTDYNAAFMS (SEQ ID NO: 262) (SEQ ID NO: 340) VH2 CDR3 NWEYDGYWGFAY (SEQ NWEYDGYWGFAY (SEQ AKNWEYDGYWGFAY (SEQ ID AKNWEYDGYWGFAY ID NO: 185) ID NO: 263) NO: 106) (SEQ ID NO: 341) VH3 CDR1 GFNIKDD (SEQ ID NO: DDYMH (SEQ ID NO: 264) GFNIKDDY (SEQ ID NO: 110) GFNIKDDYMH  186) (SEQ ID NO: 342) VH3 CDR2 DPANGN (SEQ ID NO: RIDPANGNTKYAPKFQD IDPANGNT (SEQ ID NO: 111) RIDPANGNTKYAPKFQD 187) (SEQ ID NO: 265) (SEQ ID NO: 343) VH3 CDR3 SYDGSLGDY (SEQ ID NO: SYDGSLGDY (SEQ ID ARSYDGSLGDY (SEQ ID NO: ARSYDGSLGDY  188) NO: 266) 112) (SEQ ID NO: 344) VK CDR1 TSSQSLVHSNGNTYLH TSSQSLVHSNGNTYLH QSLVHSNGNTY (SEQ ID NO: TSSQSLVHSNGNTYLH  (SEQ ID NO: 189) (SEQ ID NO: 267) 57) (SEQ ID NO: 345) VK CDR2 KVSDRFS (SEQ ID NO: KVSDRFS (SEQ ID NO: KVS (SEQ ID NO: 58) KVSDRFS (SEQ ID  190) 268) NO: 346) VK CDR3 SQSSLVPWT (SEQ ID NO: SQSSLVPWT (SEQ ID SQSSLVPWT (SEQ ID NO: 59) SQSSLVPWT (SEQ  191) NO: 269) ID NO: 347) 131E8 VH1 CDR1 GFSLPNY (SEQ ID NO: NYGVH (SEQ ID NO: 270) GFSLPNYG (SEQ ID NO: 60) GFSLPNYGVH (SEQ  192) ID NO: 348) VH1 CDR2 WSGGS (SEQ ID NO: 193) VIWSGGSTDYNAAFKS IWSGGST (SEQ ID NO: 61) VIWSGGSTDYNAAFKS (SEQ ID NO: 271) (SEQ ID NO: 349) VH1 CDR3 NFYSKYDYAMDY (SEQ ID  NFYSKYDYAMDY (SEQ ARNFYSKYDYAMDY (SEQ ID ARNFYSKYDYAMDY NO: 194) ID NO: 272) NO: 62) (SEQ ID NO: 350) VH2 CDR1 GYTFTSY (SEQ ID NO: SYWMH (SEQ ID NO: 273) GYTFTSYW (SEQ ID NO: 63) GYTFTSYWMH (SEQ  195) ID NO: 351) VH2 CDR2 NPNSGS (SEQ ID NO: NINPNSGSTNYNEKFKG INPNSGST (SEQ ID NO: 64) NINPNSGSTNYNEKFKG 196) (SEQ ID NO: 274) (SEQ ID NO: 352) VH2 CDR3 LGDY (SEQ ID NO: 197) LGDY (SEQ ID NO: 275) ARLGDY (SEQ ID NO: 65) ARLGDY (SEQ ID  NO: 353) VH3 CDR1 GFSLTSY (SEQ ID NO: SYGVH (SEQ ID NO: 276) GFSLTSYG (SEQ ID NO: 122) GFSLTSYGVH (SEQ  198) ID NO: 354) VH3 CDR2 WAGGS (SEQ ID NO: 199) VIWAGGSTNYNSALMS IWAGGST (SEQ ID NO: 123) VIWAGGSTNYNSALMS (SEQ ID NO: 277) (SEQ ID NO: 355) VH3 CDR3 DSNYFDY (SEQ ID NO: DSNYFDY (SEQ ID NO: ARDSNYFDY (SEQ ID NO: ARDSNYFDY (SEQ  200) 278) 124) ID NO: 356) VK CDR1 SASSSVSYMH (SEQ ID SASSSVSYMH (SEQ ID SSVSY (SEQ ID NO: 66) SASSSVSYMH (SEQ  NO: 201) NO: 279) ID NO: 357) VK CDR2 DTSKLAS (SEQ ID NO: DTSKLAS (SEQ ID NO: DTS (SEQ ID NO: 67) DTSKLAS (SEQ  202) 280) ID NO: 358) VK CDR3 FQGSGYPLT (SEQ ID NO: FQGSGYPLT (SEQ ID FQGSGYPLT (SEQ ID NO: 68) FQGSGYPLT (SEQ  203) NO: 281) ID NO: 359) 131H1 VH CDR1 GFSLTSY (SEQ ID NO: SYGVH (SEQ ID NO: 282)  GFSLTSYG (SEQ ID NO: 130) GFSLTSYGVH  204) (SEQ ID NO: 360) VH CDR2 WSGGS (SEQ ID NO: 205) VIWSGGSTDYNAAFIS IWSGGST (SEQ ID NO: 131) VIWSGGSTDYNAAFIS (SEQ ID NO: 283) (SEQ ID NO: 361) VH CDR3 SYDYDGRGYFDY (SEQ ID SYDYDGRGYFDY (SEQ ARSYDYDGRGYFDY (SEQ ID ARSYDYDGRGYFDY NO: 206) ID NO: 284) NO: 132) (SEQ ID NO: 362) VK1 CDR1 RASENVYRYLA (SEQ ID RASENVYRYLA (SEQ ID ENVYRY (SEQ ID NO: 136) RASENVYRYLA (SEQ NO: 207) NO: 285) ID NO: 363) VK1 CDR2 SAKTLAE (SEQ ID NO: SAKTLAE (SEQ ID NO: SAK (SEQ ID NO: 137) SAKTLAE (SEQ  208) 286) ID NO: 364) VK1 CDR3 QHHYNTPLT (SEQ ID NO: QHHYNTPLT (SEQ ID QHHYNTPLT (SEQ ID NO: 138)  QHHYNTPLT (SEQ  209) NO: 287) ID NO: 365) VK2 CDR1 KSSQSLFNSKTRKNYLA KSSQSLFNSKTRKNYLA QSLFNSKTRKNY (SEQ ID NO: KSSQSLFNSKTRKNYLA  (SEQ ID NO: 210) (SEQ ID NO: 288) 142) (SEQ ID NO: 366) VK2 CDR2 WASTRKS (SEQ ID NO: WASTRKS (SEQ ID NO: WAS (SEQ ID NO: 143) WASTRKS (SEQ  211) 289) ID NO: 367) VK2 CDR3 KQSYNLWT (SEQ ID NO: KQSYNLWT (SEQ ID NO: KQSYNLWT (SEQ ID NO: 144) KQSYNLWT (SEQ  212) 290) ID NO: 368) 132H4 VH CDR1 GFTFSNY (SEQ ID NO: NYAMS (SEQ ID NO: 291)  GFTFSNYA (SEQ ID NO: 69) GFTFSNYAMS (SEQ  213) ID NO: 369) VH CDR2 SSGGAN (SEQ ID NO: TISSGGANIYYPDSVKG ISSGGANI (SEQ ID NO: 70) TISSGGANIYYPDSVKG 214) (SEQ ID NO: 292) (SEQ ID NO: 370) VH CDR3 GDYFNHFWFAY (SEQ ID GDYFNHFWFAY (SEQ ID ARGDYFNHFWFAY (SEQ ID ARGDYFNHFWFAY  NO: 215) NO: 293) NO: 71) (SEQ ID NO: 371) VK CDR1 RSSQSIVHSNGNTYLE RSSQSIVHSNGNTYLE QSIVHSNGNTY (SEQ ID NO: RSSQSIVHSNGNTYLE  (SEQ ID NO: 216) (SEQ ID NO: 294) 72) (SEQ ID NO: 372) VK CDR2 KVSNRFS (SEQ ID NO: KVSNRFS (SEQ ID NO: KVS (SEQ ID NO: 73) KVSNRFS (SEQ ID  217) 295) NO: 373) VK CDR3 FQGSHVPWT (SEQ ID NO: FQGSHVPWT (SEQ ID FQGSHVPWT (SEQ ID NO: 74) FQGSHVPWT (SEQ  218) NO: 296) ID NO: 374) 133A6 VH CDR1 GFTFSNY (SEQ ID NO: NYAMS (SEQ ID NO: 297) GFTFSNYA (SEQ ID NO: 75) GFTFSNYAMS (SEQ  219) ID NO: 375) VH CDR2 SSGGGN (SEQ ID NO: TISSGGGNIYYTDSVKG ISSGGGNI (SEQ ID NO: 76) TISSGGGNIYYTDSVKG 220) (SEQ ID NO: 298) (SEQ ID NO: 376) VH CDR3 GDYSNYFWFAY (SEQ ID GDYSNYFWFAY (SEQ ID ARGDYSNYFWFAY (SEQ ID ARGDYSNYFWFAY  NO: 221) NO: 299) NO: 77) (SEQ ID NO: 377) VKCDR1 RSSQSIVHSNGNTYLE RSSQSIVHSNGNTYLE QSIVHSNGNTY (SEQ ID NO: RSSQSIVHSNGNTYLE  (SEQ ID NO: 222) (SEQ ID NO: 300) 78) (SEQ ID NO: 378) VK CDR2 KVSNRFS (SEQ ID NO: KVSNRFS (SEQ ID NO: KVS (SEQ ID NO: 79) KVSNRFS (SEQ  223) 301) ID NO: 379) VK CDR3 FQGSHVPWT (SEQ ID NO: FQGSHVPWT (SEQ ID FQGSHVPWT (SEQ ID NO: 80) FQGSHVPWT (SEQ  224) NO: 302) ID NO: 380) 131B4-2 VH CDR1 GFKIKDT (SEQ ID NO: DTYIH (SEQ ID NO: 303) GFKIKDTY (SEQ ID NO: 54) GFKIKDTYIH (SEQ 225) ID NO: 381) VH CDR2 DPANGN (SEQ ID NO: RIDPANGNTIYGSKFQG IDPANGNT (SEQ ID NO: 55) RIDPANGNTIYGSKFQG 226) (SEQ ID NO: 304) (SEQ ID NO: 382) VH CDR3 YVWFAY (SEQ ID NO: YVWFAY (SEQ ID NO: AGYVWFAY (SEQ ID NO: 56) AGYVWFAY (SEQ  227) 305) ID NO: 383) VK CDR1 TSSQSLVHSNGNTYLH TSSQSLVHSNGNTYLH QSLVHSNGNTY (SEQ ID NO: TSSQSLVHSNGNTYLH  (SEQ ID NO: 228) (SEQ ID NO: 306) 57) (SEQ ID NO: 384) VK CDR2 KVSDRFS (SEQ ID NO: KVSDRFS (SEQ ID NO: KVS (SEQ ID NO: 58) KVSDRFS (SEQ  229) 307) ID NO: 385) VK CDR3 SQSSLVPWT (SEQ ID NO: SQSSLVPWT (SEQ ID SQSSLVPWT (SEQ ID NO: 59) SQSSLVPWT  230) NO: 308) (SEQ ID NO: 386)

(509) TABLE-US-00011 TABLE 9  Amino Acid Sequence  Amino Acid Sequence  Antibody of Variable Region of Variable Domain DNA Sequence of Variable Region 107C6 VH MGWVVVTLPFLMAAAQSIQAQIQLVQSG QIQLVQSGPELKKPGETVKLSCRASGYT ATGGGTTGGGTGTGGACCTTGCCATTCCTGAT PELKKPGETVKLSCRASGYTFTNYGMN FTNYGMNWVKQAPGKGLKWMGWINTY GGCAGCTGCCCAAAGTATCCAAGCACAGATC WVKQAPGKGLKWMGWINTYSGVPTYA SGVPTYADDFKGQFAFSLETSAATAFLQ CAGTTGGTGCAGTCTGGTCCTGAACTGAAGA DDFKGQFAFSLETSAATAFLQINNLKDE INNLKDEDTATYFCAREGYSTTRSMDY AGCCTGGAGAGACAGTCAAGCTCTCCTGCAG DTATYFCAREGYSTTRSMDYWGQGTS WGQGTSVTVSS (SEQ ID NO 9) GGCTTCTGGATATACATTCACAAACTATGGAA VTVSSAKTTPPSVYPLA (SEQ ID  TGAACTGGGTGAAGCAGGCTCCAGGAAAGGG NO 82) TTTAAAGTGGATGGGCTGGATAAACACCTACT CTGGAGTGCCAACATATGCTGATGACTTCAAG GGACAGTTTGCCTTCTCTTTGGAAACCTCTGC CGCCACTGCCTTTTTGCAGATCAACAACCTCA AAGATGAGGACACGGCTACATATTTTTGTGCA AGAGAGGGATATAGTACTACCAGGTCTATGGA CTACTGGGGTCAAGGAACCTCAGTCACCGTC TCCTCAGCCAAAACGACACCCCCATCTGTCTA TCCACTGGCC (SEQ ID NO 81) VK M ESQSQVLILLLLWVSGTCGDIVMSQSP DIVMSQSPSSLAVSAGEKVTMSCKSSQ ATGGAGTCACAGTCTCAGGTTCTTATATTGCT SSLAVSAGEKVTMSCKSSQSLLDSRTR SLLDSRTRKNYLVWYQQKPGQSPKLLIY GCTGCTATGGGTATCTGGTACCTGTGGGGAC KNYLVWYQQKPGQSPKLLIYWASTRGS WASTRGSGVPDRFTGSGSGTDFTLTIS ATTGTGATGTCACAGTCTCCATCCTCCCTGGC GVPDRFTGSGSGTDFTLTISSVQAEDLA SVQAEDLAVYYCKQSYNLRTFGGGTKL TGTGTCAGCAGGAGAGAAGGTCACTATGAGC VYYCKQSYNLRTFGGGTKLEIKRADAAP EIK (SEQ ID NO 10) TGCAAATCCAGTCAGAGTCTGCTCGACAGTAG TVSIFPPSSEQLTSGGASVVCFLNNFYP AACCCGAAAGAACTACTTGGTTTGGTACCAGC K (SEQ ID NO 84) AGAAACCAGGGCAGTCTCCTAAACTGCTGATC TACTGGGCATCCACTAGGGGATCTGGGGTCC CTGATCGCTTCACAGGCAGTGGATCTGGGAC AGATTTCACTCTCACCATCAGCAGTGTGCAGG CTGAAGACCTGGCAGTTTATTACTGCAAACAA TCTTATAATCTTCGGACGTTCGGTGGAGGCAC CAAGCTGGAAATCAAACGGGCTGATGCTGCA CCAACTGTATCCATCTTCCCACCATCCAGTGA GCAGTTAACATCTGGAGGTGCCTCAGTCGTG TGCTTCTTGAACAACTTCTACCCCAAA (SEQ ID NO 83) 108F8 VH MGWVVVTLLFLMAAAQSIQSQIQLVQSG QIQLVQSGPDSKKPGETVKLSCRASGY ATGGGTTGGGTGTGGACCTTGCTATTCCTGAT PDSKKPGETVKLSCRASGYTFTNYGMN TFTNYGMN WVKQAPGKGLKWMGWINT GGCAGCTGCCCAAAGTATCCAATCACAGATC WVKQAPGKGLKWMGWINTYSGVPTYA YSGVPTYADDFKGQFAFSLETSAATAFL CAGTTGGTGCAGTCTGGTCCTGATTCGAAGAA DDFKGQFAFSLETSAATAFLQINNLKDE QINNLKDEDTATYFCAREGYSTTRSMDY GCCTGGAGAGACAGTCAAGCTCTCCTGCAGG DTATYFCAREGYSTTRSMDYWGQGTS WGQGTSVTVSS (SEQ ID NO 11) GCTTCTGGATATACATTCACAAACTATGGAAT VTVSSAKTTPPSVFPLAP (SEQ ID  GAACTGGGTGAAGCAGGCTCCAGGAAAGGGT NO 86) TTAAAGTGGATGGGCTGGATAAACACCTACTC TGGAGTGCCAACATATGCTGATGACTTCAAGG GACAGTTTGCCTTCTCTTTGGAAACCTCTGCC GCCACTGCCTTTTTGCAGATCAACAACCTCAA AGATGAGGACACGGCTACATATTTTTGTGCAA GAGAGGGATATAGTACTACCAGGTCTATGGA CTACTGGGGTCAAGGAACCTCAGTCACCGTC TCCTCAGCCAAAACGACACCCCCATCTGTCTT CCCCCTGGCACCT (SEQ ID NO 85) VK MGFKMKSVDLVLILLLLWVSGTCGDIVM DIVMSQSPSSLAVSAGEKVTMSCKSSQ ATGGGCTTCAAGATGAAGTCAGTCGACCTGG SQSPSSLAVSAGEKVTMSCKSSQSLLD SLLDSRTRKNYLVWYQQKPGQSPKLLIY TTCTTATATTGCTGCTGCTATGGGTATCTGGT SRTRKNYLVWYQQKPGQSPKLLIYWAS WASTRGSGVPDRFTGSGSGTDFTLTIS ACCTGTGGGGACATTGTGATGTCACAGTCTCC SVQAEDLAVYYCKQSYNLRTFGGGTKL TRGSGVPDRFTGSGSGTDFTLTISSVQA ATCCTCCCTGGCTGTGTCAGCAGGAGAGAAG EDLAVYYCKQSYNLRTFGGGTKLEIKRA EIK (SEQ ID NO 12) GTCACTATGAGCTGCAAATCCAGTCAGAGTCT DAAPTVSIFPPSSEQLTSGGASVVCFLN GCTCGACAGTAGAACCCGAAAGAACTACTTG NFYP (SEQ ID NO 88) GTTTGGTACCAGCAGAAACCAGGGCAGTCTC CTAAACTGCTGATCTACTGGGCATCCACTAGG GGATCTGGGGTCCCTGATCGCTTCACAGGCA GTGGATCTGGGACAGATTTCACTCTCACCATC AGCAGTGTGCAGGCTGAAGACCTGGCAGTTT ATTACTGCAAACAATCTTATAATCTTCGGACGT TCGGTGGAGGCACCAAGCTGGAAATCAAACG GGCTGATGCTGCACCAACTGTATCCATCTTCC CACCATCCAGTGAGCAGTTAACATCTGGAGGT GCCTCAGTCGTGTGCTTCTTGAACAACTTCTA CCCC (SEQ ID NO 87) 109A6 VH MKCSWIMFFLMAVVTGVNSEVQLQQSG EVQLQQSGAELVKPGASVKLSCTASGF ATGAAATGCAGCTGGATTATGTTCTTCCTGAT AELVKPGASVKLSCTASGFKIKDTYIHW KIKDTYIHWVIQRPAQGLEWIGRIDPANG GGCAGTGGTTACAGGGGTCAATTCAGAGGTT VIQRPAQGLEWIGRIDPANGNTIYGSKF NTIYGSKFQGKATLTADTSSNTAYIHLSS CAGCTGCAGCAGTCTGGGGCAGAACTTGTGA QGKATLTADTSSNTAYIHLSSLTSGDSA LTSGDSAVYYCAGYVWFAYWGQGTLV AGCCAGGGGCCTCAGTCAAGTTGTCCTGCAC VYYCAGYVWFAYWGQGTLVTVSAATTT TVSA (SEQ ID NO 13) AGCTTCTGGCTTCAAAATTAAAGACACCTATAT APSVFPLAP (SEQ ID NO 90) ACACTGGGTGATCCAGAGGCCTGCACAGGGC CTGGAATGGATTGGAAGGATTGATCCTGCGA ATGGTAATACTATTTATGGCTCAAAGTTCCAG GGCAAGGCCACTCTAACAGCGGACACATCAT CCAACACAGCCTACATTCACCTCAGCAGCCTG ACATCTGGGGACTCTGCCGTCTATTACTGTGC GGGCTACGTTTGGTTTGCTTACTGGGGCCAA GGGACTCTGGTCACTGTCTCTGCAGCTACAA CAACAGCCCCATCCGTCTTCCCCCTGGCACC A (SEQ ID NO 89) VK MKLPVRLLVLMFWIPASSSDVVMTQVPL DVVMTQVPLSLPVSLGDQASISCRSSQ ATGAAGTTGCCTGTTAGGCTGTTGGTGCTGAT SLPVSLGDQASISCRSSQRLVHSNGNTY RLVHSNGNTYLHWFLQKPGQSPKLLIYT GTTCTGGATTCCTGCCTCCAGCAGTGATGTTG LHWFLQKPGQSPKLLIYTVSNRFSGVPD VSNRFSGVPDRFSGSGSGTDFTLKISRV TGATGACCCAAGTTCCACTCTCCCTGCCTGTC RFSGSGSGTDFTLKISRVEAEDLGVYFC EAEDLGVYFCSQSTLVPWTFGGGTKLEI AGTCTTGGAGATCAAGCCTCCATCTCTTGCAG SQSTLVPWTFGGGTKLEIKRADAAPTVS K (SEQ ID NO 14) ATCTAGTCAGAGACTTGTGCACAGTAATGGAA IFPPSSEQLTSGGASVVCFLNNFYPK ACACCTATTTACATTGGTTCTTACAGAAGCCA (SEQ ID NO 92) GGCCAGTCTCCAAAGCTCCTGATCTACACAGT TTCCAACCGATTTTCTGGGGTCCCAGACAGGT TCAGTGGCAGTGGATCAGGGACAGATTTCAC ACTCAAGATCAGCAGAGTGGAGGCTGAGGAT CTGGGAGTTTATTTCTGCTCTCAAAGTACACT TGTTCCGTGGACGTTCGGTGGAGGCACCAAG CTGGAAATCAAACGGGCTGATGCTGCACCAA CTGTATCCATCTTCCCACCATCCAGTGAGCAG TTAACATCTGGAGGTGCCTCAGTCGTGTGCTT CTTGAACAACTTCTACCCAAAG (SEQ ID  NO 91) 111A6 VH MKCSWVMFFLMAVVTGVNSEVQLQQS EVQLQQSGAELVKPGASVKLSCTASGF ATGAAATGCAGCTGGGTTATGTTCTTCCTGAT GAELVKPGASVKLSCTASGFKIKDTYIH KIKDTYIHWVIQRPAQGLEWIGRIDPANG GGCAGTGGTTACAGGGGTCAATTCAGAGGTT WVIQRPAQGLEWIGRIDPANGNTIYGSK NTIYGSKFQGKATLTADTSSNTAYIHLSS CAGCTGCAGCAGTCTGGGGCAGAACTTGTGA FQGKATLTADTSSNTAYIHLSSLTSGDS LTSGDSAVYYCAGYVWFAYWGQGTLV AGCCAGGGGCCTCAGTCAAGTTGTCCTGCAC AVYYCAGYVWFAYWGQGTLVTVSAATT TVSA (SEQ ID NO 15) AGCTTCTGGCTTCAAAATTAAAGACACCTATAT TAPSVFPLAP (SEQ ID NO 94) ACACTGGGTGATCCAGAGGCCTGCACAGGGC CTGGAATGGATTGGAAGGATTGATCCTGCGA ATGGTAATACTATTTATGGCTCAAAGTTCCAG GGCAAGGCCACTCTAACAGCGGACACATCAT CCAACACAGCCTACATTCACCTCAGCAGCCTG ACATCTGGGGACTCTGCCGTCTATTACTGTGC GGGCTACGTTTGGTTTGCTTACTGGGGCCAA GGGACTCTGGTCACTGTCTCTGCAGCTACAA CAACAGCCCCATCCGTCTTCCCCCTGGCACC A (SEQ ID NO 93) VK1 MDFQVQIFSFLLISASVAMSRGENVLTQ ENVLTQSPAIMSASPGEKVTMTCRARS ATGGATTTTCAGGTGCAGATTTTCAGCTTCTT SPAIMSASPGEKVTMTCRARSSVSSSYL SVSSSYLHWYQQKSGASPKLWIYSTSN GCTAATCAGTGCCTCAGTTGCAATGTCCAGAG HWYQQKSGASPKLWIYSTSNLASGVPT LASGVPTRFSGSGSGTSYSLTISSVEAE GAGAAAATGTGCTCACCCAGTCTCCAGCAATC RFSGSGSGTSYSLTISSVEAEDAATYYC DAATYYCQQYSGYPLTFGAGTKLELK ATGTCTGCTTCTCCAGGGGAGAAGGTCACCA QQYSGYPLTFGAGTKLELKRADAAPTV (SEQ ID NO 16) TGACCTGCAGGGCCAGGTCAAGTGTAAGTTC SIFPPSSEQLTSGGASVVCFLNNFYPK CAGTTACTTGCACTGGTACCAGCAGAAGTCAG (SEQ ID NO 96) GTGCCTCCCCCAAACTCTGGATTTATAGCACA TCCAACTTGGCTTCTGGAGTCCCTACTCGCTT CAGTGGCAGTGGGTCTGGGACCTCTTACTCT CTCACAATCAGCAGTGTGGAGGCTGAAGATG CTGCCACTTATTACTGCCAGCAGTACAGTGGT TACCCACTCACGTTCGGTGCTGGGACCAAGC TGGAGCTGAAACGGGCTGATGCTGCACCAAC TGTATCCATCTTCCCACCATCCAGTGAGCAGT TAACATCTGGAGGTGCCTCAGTCGTGTGCTTC TTGAACAACTTCTACCCCAAG (SEQ ID  NO 95) VK2 MKLPVRLLVLMFWIPASSSDVVMTQVPL DVVMTQVPLSLPVSLGDQASISCRSSQ ATGAAGTTGCCTGTTAGGCTGTTGGTGCTGAT SLPVSLGDQASISCRSSQRLVHSNGNTY RLVHSNGNTYLHWFLQKPGQSPKLLIYT GTTCTGGATTCCTGCCTCCAGCAGTGATGTTG LHWFLQKPGQSPKLLIYTVSNRFSGVPD VSNRFSGVPDRFSGSGSGTDFTLKISRV TGATGACCCAAGTTCCACTCTCCCTGCCTGTC RFSGSGSGTDFTLKISRVEAEDLGVYFC EAE DLGVYFCSQSTLVPWTFGGGTKLEI AGTCTTGGAGATCAAGCCTCCATCTCTTGCAG SQSTLVPWTFGGGTKLEIKRADAAPTVS K (SEQ ID NO 17) ATCTAGTCAGAGACTTGTGCACAGTAATGGAA IFPPSSEQLTSGGASVVCFLN NFYPK ACACCTATTTACATTGGTTCTTACAGAAGCCA (SEQ ID NO 98) GGCCAGTCTCCAAAGCTCCTGATCTACACAGT TTCCAACCGATTTTCTGGGGTCCCAGACAGGT TCAGTGGCAGTGGATCAGGGACAGATTTCAC ACTCAAGATCAGCAGAGTGGAGGCTGAGGAT CTGGGAGTTTATTTCTGCTCTCAAAGTACACT TGTTCCGTGGACGTTCGGTGGAGGCACCAAG CTGGAAATCAAACGGGCTGATGCTGCACCAA CTGTATCCATCTTCCCACCATCCAGTGAGCAG TTAACATCTGGAGGTGCCTCAGTCGTGTGCTT CTTGAACAACTTCTACCCCAAAG (SEQ  ID NO 97) 131B4 VH1 MKCSWIMFFLMAVVTGVNSEVQVQQS EVQVQQSGAELVKPGASVKLSCTASGF ATGAAATGCAGCTGGATTATGTTCTTCCTGAT GAELVKPGASVKLSCTASGFKIKDTYIH KIKDTYIHWLKQRPEQGLEWIGRIDPAN GGCAGTGGTTACAGGGGTCAATTCAGAGGTT WLKQRPEQGLEWIGRIDPANGNTIYGSK GNTIYGSKFQGKATITADTSSNTAYIQLS CAGGTGCAGCAGTCTGGGGCAGAGCTTGTGA FQGKATITADTSSNTAYIQLSSLTSGDTA SLTSGDTAVYFCAGYVWFAYWGQGTLV AGCCAGGGGCCTCAGTCAAGTTGTCCTGCAC VYFCAGYVWFAYWGQGTLVTVSAAKTT TVSA (SEQ ID NO 18) AGCTTCTGGCTTCAAAATTAAGGACACCTATA PPSVFPLA (SEQ ID NO 100) TACACTGGTTAAAACAGAGGCCTGAACAGGG CCTGGAATGGATTGGAAGGATTGATCCTGCG AATGGTAATACTATATATGGCTCAAAGTTCCA GGGCAAGGCCACTATAACAGCAGACACATCA TCCAACACAGCCTACATTCAACTCAGCAGCCT GACATCTGGGGACACTGCCGTCTATTTTTGTG CGGGCTACGTTTGGTTTGCTTACTGGGGCCA AGGGACTCTGGTCACTGTCTCTGCAGCCAAA ACGACACCCCCATCCGTCTTCCCCCTGGCC (SEQ ID NO 99) VH2 MAVLGLLFCLVTFPSCVLSQVQLKQSGP QVQLKQSGPSLVQPSQSLSITCTVSGFS ATGGCTGTCTTGGGGCTGCTCTTCTGCCTGG SLVQPSQSLSITCTVSGFSLTSYGVHWV LTSYGVHWVRQSPGKGLEWLGVIWRG TGACATTCCCAAGCTGTGTCCTGTCCCAGGTG RQSPGKGLEWLGVIWRGGSTDYNAAF GSTDYNAAFMSRLSITKDNSKSQVFFKM CAGCTGAAGCAGTCAGGACCTAGCCTAGTGC MSRLSITKDNSKSQVFFKMNSLQADDTA NSLQADDTAIYYCAKNWEYDGYWGFAY AGCCCTCACAGAGCCTGTCCATAACCTGCAC IYYCAKNWEYDGYWGFAYWGQGTLVT WGQGTLVTVSA (SEQ ID NO 103) AGTCTCTGGTTTCTCATTAACTAGCTATGGTG VSAESQSFPNVFPLE (SEQ ID NO  TACACTGGGTTCGCCAGTCTCCAGGAAAGGG 102) TCTGGAGTGGCTGGGAGTGATATGGAGAGGT GGAAGCACAGACTACAATGCAGCTTTCATGTC CAGACTGAGCATCACCAAGGACAACTCCAAG AGCCAAGTTTTCTTTAAAATGAACAGTCTGCA AGCTGATGACACTGCCATATACTACTGTGCCA AAAATTGGGAGTATGATGGTTACTGGGGGTTT GCTTACTGGGGCCAAGGGACTCTGGTCACTG TCTCTGCAGAGAGTCAGTCCTTCCCAAATGTC TTCCCCCTCGAA (SEQ ID NO 101) VH3 MAVVTGVNSEVQLQQSGAELVRPGASV EVQLQQSGAELVRPGASVKLSCTASGF ATGGCAGTGGTTACAGGGGTCAATTCAGAGG KLSCTASGFNIKDDYMHWVKQRPEQGL NIKDDYMHWVKQRPEQGLEWIGRIDPA TTCAGCTGCAGCAGTCTGGGGCTGAGCTTGT EWIGRIDPANGNTKYAPKFQDKATITAD NGNTKYAPKFQDKATITADTSSNTAYLQ GAGGCCAGGGGCCTCAGTCAAGTTGTCCTGC TSSNTAYLQLSSLTSEDTAVYYCARSYD LSSLTSEDTAVYYCARSYDGSLGDYWG ACAGCTTCTGGCTTTAACATTAAAGACGACTA GSLGDYWGQGTTLTVSSESQSFPNVFP QGTTLTVSS (SEQ ID NO 109) TATGCACTGGGTGAAGCAGAGGCCTGAACAG LE (SEQ ID NO 108) GGCCTGGAGTGGATTGGAAGGATTGATCCTG CGAATGGTAATACTAAATATGCCCCGAAGTTC CAGGACAAGGCCACTATAACTGCAGACACAT CCTCCAACACAGCCTACCTGCAGCTCAGCAG CCTGACATCTGAGGACACTGCCGTCTATTACT GTGCTAGAAGCTATGATGGTTCTCTGGGGGA CTACTGGGGCCAAGGCACCACTCTCACAGTC TCCTCAGAGAGTCAGTCCTTCCCAAATGTCTT CCCCCTCGAG (SEQ ID NO 107) VK MKLPVRLLVLMFWIPASSSDAVLTQTPL DAVLTQTPLSLPVSLGDQASISCTSSQS ATGAAGTTGCCTGTTAGGCTGTTGGTGCTGAT SLPVSLGDQASISCTSSQSLVHSNGNTY LVHSNGNTYLHWYLQKPGQSPKLLIYKV GTTCTGGATTCCTGCTTCCAGCAGTGATGCTG LHWYLQKPGQSPKLLIYKVSDRFSGVPD SDRFSGVPDRFSGSGSGTDFTLMITRV TGTTGACCCAAACTCCACTCTCCCTGCCTGTC RFSGSGSGTDFTLMITRVEAEDLGVYFC EAEDLGVYFCSQSSLVPWTFGGGTKLE AGTCTTGGAGATCAAGCCTCCATCTCTTGCAC SQSSLVPWTFGGGTKLEVKRADAAPTV VK (SEQ ID NO 19) ATCTAGTCAGAGCCTTGTACACAGTAATGGAA SIFPPSSEQLTSGGASVVCFLNNFYPK ACACCTATTTACATTGGTACCTGCAGAAGCCA (SEQ ID NO 114) GGCCAGTCTCCAAAGCTCCTGATCTACAAAGT TTCCGACCGATTTTCTGGGGTCCCAGACAGG TTCAGTGGCAGTGGATCAGGAACAGATTTCAC ACTCATGATCACCAGAGTGGAGGCTGAGGAT CTGGGAGTTTATTTCTGCTCTCAAAGTTCACTT GTTCCGTGGACGTTCGGTGGAGGCACCAAGC TGGAAGTCAAACGGGCTGATGCTGCACCAAC TGTATCCATCTTCCCACCATCCAGTGAGCAGT TAACATCTGGAGGTGCCTCAGTCGTGTGCTTC TTGAACAACTTCTACCCCAAA (SEQ ID NO 113) 131E8 VH1 MAVLGLLFCLVTFPSCVLSQVQLKQSRP QVQLKQSRPGPVQPSQSLSITCTVSGF ATGGCTGTTTTGGGGCTGCTCTTCTGCCTGGT GPVQPSQSLSITCTVSGFSLPN YGVHW SLPNYGVHWVRQPPGKGLEWLGVIWS GACATTCCCAAGCTGTGTCCTATCCCAGGTGC VRQPPGKGLEWLGVIWSGGSTDYNAAF GGSTDYNAAFKSRLSISKDNSKSQVFFK AGCTGAAGCAGTCAAGACCTGGCCCAGTGCA KSRLSISKDNSKSQVFFKMNSLQADDTA MNSLQADDTAIYYCARNFYSKYDYAMD GCCCTCACAGAGCCTGTCCATCACCTGCACA IYYCARNFYSKYDYAMDYWGQGTSVTV YWGQGTSVTVSS (SEQ ID NO 20) GTCTCTGGTTTCTCATTACCTAACTATGGTGTA SSAKTTPPSVFPL (SEQ ID NO 116) CACTGGGTTCGCCAGCCTCCAGGAAAGGGTC TGGAGTGGCTGGGAGTGATATGGAGTGGTGG AAGCACAGACTATAATGCAGCTTTCAAATCCA GACTGAGCATCAGCAAGGACAACTCCAAGAG CCAAGTTTTCTTTAAAATGAACAGTCTGCAAG CTGATGACACAGCCATATACTACTGTGCCAGA AATTTTTATAGTAAGTACGACTATGCTATGGAC TACTGGGGTCAAGGAACCTCAGTCACCGTCT CCTCAGCCAAAACAACACCCCCATCCGTCTTC CCCCTGGC (SEQ ID NO 115) VH2 MFFLVATATGVHSQVQLQQPGSVLVRP QVQLQQPGSVLVRPGASVKLSCKASGY ATGTTCTTCTTGGTAGCAACAGCTACAGGTGT GASVKLSCKASGYTFTSYWMHWVKQR TFTSYWMHWVKQRPGQGLEWIGNIINP CCACTCCCAGGTCCAACTGCAGCAGCCTGGG PGQGLEWIGNINPNSGSTNYNEKFKGK NSGSTNYNEKFKGKATLTVDTSSSTAY TCTGTGCTGGTGAGGCCTGGAGCTTCAGTGA ATLTVDTSSSTAYMDLSSLTSEDSAVYY MDLSSLTSEDSAVYYCARLGDYWGQGT AGCTGTCCTGCAAGGCTTCTGGCTACACATTC CARLGDYWGQGTTLTVSSKSQSSPSVF TLTVSS (SEQ ID NO 21) ACCAGCTACTGGATGCACTGGGTGAAGCAGA PL (SEQ ID NO 118) GGCCGGGACAAGGCCTTGAGTGGATTGGAAA TATTAATCCTAATAGTGGTAGTACTAACTACAA TGAGAAGTTCAAGGGCAAGGCCACACTGACT GTAGACACATCCTCCAGCACAGCCTACATGG ATCTCAGCAGCCTGACATCTGAGGACTCTGC GGTCTATTACTGTGCAAGACTGGGTGACTACT GGGGCCAAGGCACCACTCTCACAGTCTCCTC AAAGAGTCAGTCCTCCCCATCCGTCTTCCCCC TG (SEQ ID NO 117) VH3 AVLGLLFCLVAFPSCVLSQVQLKESGPG QVQLKESGPGLVAPSQSLSITCTVSGFS GCTGTCTTGGGGCTGCTCTTCTGCCTGGTTG LVAPSQSLSITCTVSGFSLTSYGVHWVR LTSYGVHWVRQPPGKGLEWLGVIWAG CATTTCCAAGCTGTGTCCTGTCCCAGGTGCAG QPPGKGLEWLGVIWAGGSTNYNSALMS GSTNYNSALMSRLSISKDNSKSQVFLKM CTGAAGGAGTCAGGACCTGGCCTGGTGGCGC RLSISKDNSKSQVFLKMNSLQTDDTAMY NSLQTDDTAMYYCARDSNYFDYWGQG CCTCACAGAGCCTGTCCATCACTTGCACTGTC YCARDSNYFDYWGQGTTLTVSSESQSF TTLTVSS (SEQ ID NO 121) TCTGGGTTTTCATTAACCAGCTATGGTGTACA PNVFPLV (SEQ ID NO 120) CTGGGTTCGCCAGCCTCCAGGAAAGGGTCTG GAGTGGCTGGGAGTAATATGGGCTGGTGGAA GCACAAATTATAATTCGGCTCTCATGTCCAGA CTGAGCATCAGCAAAGACAACTCCAAGAGCC AAGTTTTCTTAAAAATGAACAGTCTGCAAACTG ATGACACAGCCATGTACTACTGTGCCAGAGAT AGTAACTACTTTGACTACTGGGGCCAAGGCAC CACTCTCACAGTCTCCTCAGAGAGTCAGTCCT TCCCAAATGTCTTCCCCCTCGTA (SEQ ID NO 119) VK MDFQVQIFSFLLISASVIMSRGENVLTQS ENVLTQSPAIMSASPGEKVTMTCSASSS ATGGATTTTCAGGTGCAGATTTTCAGCTTCCT PAIMSASPGEKVTMTCSASSSVSYMHW VSYMHWYQQKSSTSPKLWIYDTSKLAS GCTAATCAGTGCCTCAGTCATAATGTCCAGAG YQQKSSTSPKLWIYDTSKLASGVPGRFS GVPGRFSGSGSGNSYSLTISSMEAEDV GAGAAAATGTTCTCACCCAGTCTCCAGCAATC GSGSGNSYSLTISSMEAEDVATYYCFQ ATYYCFQGSGYPLTFGSGTKLEIK ATGTCTGCATCTCCAGGGGAAAAGGTCACCAT GSGYPLTFGSGTKLEIKRADAAPTVSIFP (SEQ ID NO 22) GACCTGCAGTGCCAGCTCAAGTGTAAGTTACA PSSEQLTSGGASVVCFLNNFYPK (SEQ TGCACTGGTACCAGCAGAAGTCAAGCACCTC ID NO 126) CCCCAAACTCTGGATTTATGACACATCCAAAC TGGCTTCTGGAGTCCCAGGTCGCTTCAGTGG CAGTGGGTCTGGAAACTCTTACTCTCTCACGA TCAGCAGCATGGAGGCTGAAGATGTTGCCAC TTATTACTGTTTTCAGGGGAGTGGGTACCCAC TCACGTTCGGCTCGGGGACAAAGTTGGAAAT AAAACGGGCTGATGCTGCACCAACTGTATCCA TCTTCCCACCATCCAGTGAGCAGTTAACATCT GGAGGTGCCTCAGTCGTGTGCTTCTTGAACA ACTTCTACCCCAAA (SEQ ID NO 125) 131H1 VH MAVLGLLFCLVTFPSCVLSQVQLKQSGP QVQLKQSGPGLVQPSQSLSITCTVSGFS ATGGCTGTCTTGGGGCTGCTCTTCTGCCTGG GLVQPSQSLSITCTVSGFSLTSYGVHWV LTSYGVHWVRQSPGKGLEWLGVIWSG TGACATTCCCAAGCTGTGTCCTATCCCAGGTG RQSPGKGLEWLGVIWSGGSTDYNAAFI GSTDYNAAFISRLSISKDNSKSQVFFKM CAGCTGAAGCAGTCAGGACCTGGCCTAGTGC SRLSISKDNSKSQVFFKMNSLQADDTAI NSLQADDTAIYYCARSYDYDGRGYFDY AGCCCTCACAGAGCCTGTCCATCACCTGCAC YYCARSYDYDGRGYFDYWGQGTTLTV WGQGTTLTVSS (SEQ ID NO 129) AGTCTCTGGTTTCTCATTAACTAGCTATGGTG SSESQSFPNVFPLV (SEQ ID NO  TACACTGGGTTCGCCAGTCTCCAGGAAAGGG 128) TCTGGAGTGGCTGGGAGTGATATGGAGTGGT GGAAGCACAGACTATAATGCAGCTTTCATATC CAGACTGAGCATCAGCAAGGACAATTCCAAG AGCCAAGTTTTCTTTAAAATGAACAGTCTGCA AGCTGATGACACAGCCATATATTACTGTGCCA GATCTTATGATTACGACGGGAGGGGTTACTTT GACTACTGGGGCCAAGGCACCACTCTCACAG TCTCCTCAGAGAGTCAGTCCTTCCCAAATGTC TTCCCCCTCGTA (SEQ ID NO 127) VK1 MSVLTQVLGLLLLWLTGARCDIQMTQSP DIQMTQSPASLSASVGETVTITCRASEN ATGAGTGTGCTCACTCAGGTCCTGGGGTTGC ASLSASVGETVTITCRASENVYRYLAWY VYRYLAWYQQRQGKSPQLLVYSAKTLA TGCTGCTGTGGCTTACAGGTGCCAGATGTGA QQRQGKSPQLLVYSAKTLAEGVPSRFS EGVPSRFSGSGSGTQFSLKINTLQPEDF CATCCAGATGACTCAGTCTCCAGCCTCCCTGT GSGSGTQFSLKINTLQPEDFGTYYCQH GTYYCQHHYNTPLTFGAGTKLELK CTGCATCTGTGGGAGAAACTGTCACCATCACA HYNTPLTFGAGTKLELKRADAAPTVSIFP  (SEQ ID NO 135) TGTCGAGCAAGTGAGAATGTTTACAGATATTT PSSEQLTSGGASVVCFLN NFYPK  AGCATGGTATCAGCAGAGACAGGGAAAATCT (SEQ ID NO 134) CCTCAGCTCCTGGTCTATAGTGCAAAAACCTT AGCAGAAGGTGTGCCATCAAGGTTCAGTGGC AGTGGATCAGGCACACAGTTTTCTCTGAAGAT CAACACCCTGCAGCCTGAAGATTTTGGGACTT ATTACTGTCAACATCATTATAATACTCCTCTCA CGTTCGGTGCTGGGACCAAGCTGGAGCTGAA ACGGGCTGATGCTGCACCAACTGTATCCATCT TCCCACCATCCAGTGAGCAGTTAACATCTGGA GGTGCCTCAGTCGTGTGCTTCTTGAACAACTT CTACCCCAAA (SEQ ID NO 133) VK2 MVLIWLLLWVSGTCGDIVMSQSPSSLAV DIVMSQSPSSLAVSAGEKVTMSCKSSQ ATGGTTCTTATATGGCTCCTGCTATGGGTATC SAGEKVTMSCKSSQSLFNSKTRKNYLA SLFNSKTRKNYLAWFQQKPGQSPELLIY TGGTACCTGTGGGGACATTGTGATGTCACAGT WFQQKPGQSPELLIYWASTRKSGVPDR WASTRKSGVPDRFTGSGSGTDFTLTISS CTCCATCCTCCCTGGCTGTGTCAGCAGGAGA FTGSGSGTDFTLTISSVQAEDLAVYYCK VQAEDLAVYYCKQSYNLWTFGGGTKLE GAAGGTCACTATGAGCTGCAAATCCAGTCAGA QSYNLWTFGGGTKLEIKRADAAPTVSIF IK (SEQ ID NO 141) GTCTGTTCAACAGTAAAACCCGAAAGAACTAC PPSSEQLTSGGASVVCFLNNFYPK TTGGCTTGGTTTCAGCAAAAACCAGGGCAGTC (SEQ ID NO 140) TCCTGAACTGCTGATCTACTGGGCATCCACTA GGAAATCTGGGGTCCCTGATCGCTTCACAGG CAGTGGATCTGGGACAGATTTCACTCTCACCA TCAGCAGTGTGCAGGCTGAAGACCTGGCAGT TTATTACTGCAAGCAATCTTATAATCTGTGGAC GTTCGGCGGAGGCACCAAGCTGGAAATCAAA CGGGCTGATGCTGCACCAACTGTATCCATCTT CCCACCATCCAGTGAGCAGTTAACATCTGGA GGTGCCTCAGTCGTGTGCTTCTTGAACAACTT CTACCCCAAA (SEQ ID NO 139) 132H4 VH SWVFLVLILKGVQCEVKLVESGGGLVKP EVKLVESGGGLVKPGGSLKLSCAASGF TGAGCTGGGTTTTCCTTGTCCTTATTTTAAAAG GGSLKLSCAASGFTFSNYAMSWVRQNP TFSNYAMSWVRQNPAKRLEWVATISSG GTGTCCAGTGTGAAGTGAAGCTGGTGGAGTC AKRLEWVATISSGGANIYYPDSVKGRFII GANIYYPDSVKGRFIISRDNARNTLYLQ TGGGGGAGGCTTAGTGAAGCCTGGAGGGTCC SRDNARNTLYLQMSSLRSEDTAMYYCA MSSLRSEDTAMYYCARGDYFNHFWFA CTGAAACTCTCCTGTGCAGCCTCTGGATTCAC RGDYFNHFWFAYWGQGTLVTVSAAKTT YWGQGTLVTVSA (SEQ ID NO 23) TTTCAGTAACTATGCCATGTCTTGGGTTCGCC APSVFPLA (SEQ ID NO 146) AGAATCCGGCGAAGAGGCTGGAGTGGGTCG CAACCATTAGTAGTGGTGGTGCTAATATTTAC TATCCAGACAGTGTGAAGGGCCGATTCATCAT CTCCAGAGACAATGCCAGGAACACCCTGTAC CTGCAAATGAGCAGTCTGAGGTCTGAGGACA CGGCCATGTATTACTGTGCAAGAGGCGACTAT TTTAACCACTTCTGGTTTGCTTACTGGGGCCA AGGGACTCTTGTCACTGTCTCTGCAGCCAAAA CAACAGCCCCATCGGTCTTCCCCCTGGCA (SEQ ID NO 145) VK MKLPVRLLVLMFWIPASSSDVLMTQTPL DVLMTQTPLSLPVSLGDQASISCRSSQS ATGAAGTTGCCTGTTAGGCTGTTGGTGCTGAT SLPVSLGDQASISCRSSQSIVHSNGNTY IVHSNGNTYLEWYLQKPGQSPKFLIYKV GTTCTGGATTCCTGCTTCCAGCAGTGATGTTT SNRFSGVPDRFSGSGSGTDFTLKINRVE LEWYLQKPGQSPKFLIYKVSNRFSGVPD TGATGACCCAAACTCCACTCTCCCTGCCTGTC RFSGSGSGTDFTLKINRVEAEDLGIYYC AEDLGIYYCFQGSHVPWTFGGGTKLEIK AGTCTTGGAGATCAAGCCTCCATCTCTTGTAG FQGSHVPVVTFGGGTKLEIKRADAAPTV RA (SEQ ID NO 24) ATCGAGTCAGAGCATTGTACATAGTAATGGAA SIFPPSSEQLTSGGASVVCFL (SEQ  ACACCTATTTAGAATGGTACCTGCAGAAACCA ID NO 148) GGCCAGTCTCCAAAGTTCCTGATCTACAAAGT TTCCAACCGATTTTCAGGGGTCCCAGACAGGT TCAGTGGCAGTGGATCAGGGACAGATTTCAC ACTCAAGATCAACAGAGTGGAGGCTGAGGAT CTGGGAATTTATTACTGCTTTCAGGGTTCACA TGTTCCGTGGACGTTCGGTGGAGGCACCAAG CTGGAAATCAAACGGGCTGATGCTGCACCAA CTGTATCCATCTTCCCACCATCCAGTGAGCAG TTAACATCTGGAGGTGCCTCAGTCGTGTGCTT CTTGA (SEQ ID NO 147) 133A6 VH MNFGLRLVFLVLVLKGVQCEVKLVESG EVKLVESGGGLVKPGGSLKLSCAASGF ATGAACTTTGGGTTGAGATTGGTTTTCCTTGT GGLVKPGGSLKLSCAASGFTFSNYAMS TFSNYAMSWVRQTPAKRLEWVTTISSG CCTTGTTTTAAAAGGTGTCCAGTGTGAGGTGA WVRQTPAKRLEWVTTISSGGGNIYYTD GGNIYYTDSVKGRFTVSRDNARNTLYLQ AGCTAGTGGAGTCTGGAGGAGGCTTAGTGAA SVKGRFTVSRDNARNTLYLQMSSLRSE MSSLRSEDTAMYYCARGDYSNYFWFA GCCTGGAGGGTCCCTGAAACTCTCCTGTGCA DTAMYYCARGDYSNYFWFAYWGQGTL YWGQGTLVSVSE (SEQ ID NO 25) GCCTCTGGATTCACTTTCAGTAACTATGCCAT VSVSEAKTTAPSVFPLAP (SEQ  GTCTTGGGTTCGCCAGACTCCGGCGAAGAGG ID NO 150) CTGGAGTGGGTCACAACCATTAGTAGTGGTG GTGGTAACATCTACTATACAGACAGTGTGAAG GGCCGATTCACCGTCTCCAGAGACAATGCCA GGAACACCCTGTACCTGCAAATGAGCAGTCT GAGGTCTGAGGACACGGCCATGTATTACTGT GCAAGAGGCGACTATAGTAACTACTTCTGGTT TGCTTACTGGGGCCAAGGGACTCTGGTCTCT GTCTCTGAAGCCAAAACAACAGCCCCATCGG TCTTCCCCCTGGCACCT (SEQ ID NO 149) VK MKLPVRLLVLMFWIPASSSDVLMTQTPL DVLMTQTPLSLPVSLGDQASISCRSSQS ATGAAGTTGCCTGTTAGGCTGTTGGTGCTGAT SLPVSLGDQASISCRSSQSIVHSNGNTY IVHSNGNTYLEWYLQKPGQSPKLLIYKV GTTCTGGATTCCTGCTTCCAGCAGTGATGTTT LEWYLQKPGQSPKLLIYKVSNRFSGVPD SNRFSGVPDRFSGSGSGTDFTLKISRVE TGATGACCCAAACTCCACTCTCCCTGCCTGTC RFSGSGSGTDFTLKISRVEAEDLGVYYC AEDLGVYYCFQGSHVPWTFGGGTKLEI AGTCTTGGAGATCAAGCCTCCATCTCTTGCAG FQGSHVPVVTFGGGTKLEIKRADAAPTV K (SEQ ID NO 26) ATCTAGTCAGAGCATTGTACATAGTAATGGAA SIFPPSREQLTSGGASVVCFLNNFYPK ACACCTATTTAGAATGGTACCTGCAGAAACCA (SEQ ID NO 152) GGCCAGTCTCCAAAGCTCCTGATCTACAAAGT TTCCAACCGATTTTCTGGGGTCCCAGACAGGT TCAGTGGCAGTGGATCAGGGACAGATTTCAC ACTCAAGATCAGCAGAGTGGAGGCTGAGGAT CTGGGAGTTTATTACTGCTTTCAAGGTTCACA TGTTCCGTGGACGTTCGGTGGAGGCACCAAG CTGGAAATCAAACGGGCTGATGCTGCACCAA CTGTATCCATCTTCCCACCATCCAGGGAGCAG TTAACATCTGGAGGTGCCTCAGTCGTGTGCTT CTTGAACAACTTCTACCCAAAA (SEQ ID NO 151) 131B4-2 VH EVQLQQSGAELVKPGASVKLSCTASGF KIKDTYIHWLKQRPEQGLEWIGRIDPAN GNTIYGSKFQGKATITADTSSNTAYIQLS SLTSGDTAVYFCAGYVWFAYWGQGTLV TVSA (SEQ ID NO 387) VK DAVLTQTPLSLPVSLGDQASISCTSSQS LVHSNGNTYLHWYLQKPGQSPKLLIYKV SDRFSGVPDRFSGSGSGTDFTLMITRV EAEDLGVYFCSQSSLVPWTFGGGTKLE VK (SEQ ID NO: 19)

REFERENCES

(510) Argiriadi M A, Xiang T, Wu C, Ghayur T and Borhani D W. Unusual water-mediated antigenic recognition of the proinflammatory cytokine interleukin-18. J Biol Chem 2009; 284(36)24478-24489. Azoulay E, Eddahibi S, Marcos E, Levame M, Harf A, Schlemmer B, Adnot S and Delclaux C. Granulocyte colony-stimulating factor enhances alpha-naphthylthiourea-induced pulmonary hypertension. J Appl Physiol 2003; 94:2027-2033. Baron R M, Choi A J S, Owen C A and Choi A M K. Genetically manipulated mouse models of lung disease: potential and pitfalls. Am J Physiol Lung Cell Mol Physiol 2012; L485-L497. Chen D Y, Lan J L, Lin F J and Hsieh T Y. Proinflammatory cytokine profiles in sera and pathological tissues of patients with active untreated adult onset Still's disease. J Rheumatol 2004; 31:2189-2198. Chen D Y, Lan J L, Lin F J, Hsieh T Y and Wen M C. Predominance of Th1 cytokine in peripheral blood and pathological tissues of patients with active untreated adult onset Still's disease. Ann Rheum Dis 2004; 63(10):1300-1306. Cunningham R E. Tissue disaggregation. Methods Mol Biol 1994; 34:225-228. Daley E, Emson C, Guignabert C, de Waal Malefyt R, Louten J, Kurup V P, Hogaboam C, Taraseviciene-Stewart L, Voelkel N F, Rabinovitch M, et al. Pulmonary arterial remodeling induced by a Th2 immune response. J Exp Med 2008; 205:361-372. Elias J A, Kang M J, Crothers K et al. Mechanistic heterogeneity in chronic obstructive pulmonary disease: insights from transgenic mice. Proc Am Thorac Soc 2006; 494-498. Eltom S, Stevenson C S and Rastrick J. P2X7 Receptor and Caspase—1 Activation Are Central to Airway Inflammation Observed after Exposure to Tobacco Smoke PLoS ONE 2011; 6(9):e24097. Hackett B P, Shimizu N and Gitlin J D. Clara cell secretory protein gene expression in bronchiolar epithelium. Am J Physiol 1992; 262:L399-L404. Halbower A C, Mason R J, Abman S H and Tuder R M. Agarose infiltration improves morphology of cryostat sections of lung. Lab Invest 1994; 71: 149-153. Hautamaki R D, Kobayashi D K, Senior R M and Shapiro S D. Requirement for macrophage elastase for cigarette smoke-induced emphysema in mice. Science 1997; 277: 2002-2004. Hoshino T, Kawase Y, Okamoto M et al. IL-18-transgenic mice: in vivo evidence of a broad role for IL-18 in modulating immune function. J Immunol 2001; 7014-7018. Hoshino T, Kato S, Oka N et al. Pulmonary inflammation and emphysema: role of the cytokines IL-18 and IL-13. Am J Respir Crit Care Med 2007; 49-62. Helmut Fenner. Targeting IL-18 in Chronic Obstructive Pulmonary Disease: Background and rationale. Mar. 22, 2013 CONFIDENTIAL Page 19. Hou S, Li B, Wang L, Qian W, Zhang D, Hong X, Wang H, Guo Y (July 2008). “Humanization of an anti-CD34 monoclonal antibody by complementarity-determining region grafting based on computer-assisted molecular modeling.”. J Biochem 144 (1): 115-20 Imaoka H, Hoshino T, Takei S, Kinoshita T et al. Interleukin-18 production and pulmonary function in COPD. Eur Respir J 2008; 287-97. Jaatinen T, Laine J. Isolation of mononuclear cells from human cord blood by Ficoll-Paque density gradient. Curr Protoc Stem Cell Biol 2007; Chapter 2:Unit 2A.1. Kang M J, Lee C G, Lee J Y, Dela Cruz C S, Chen Z J, Enelow R, Elias J A. Cigarette smoke selectively enhances viral PAMP- and virus-induced pulmonary innate immune and remodeling responses in mice. J Clin Invest 2008; 118:2771-2784. Kang M J, Homer R J, Gallo A et al. IL-18 is induced and IL-18 receptor alpha plays a critical role in the pathogenesis of cigarette smoke-induced pulmonary emphysema and inflammation. J Immunol 2007; 1948-1959. Kang M J, Choi J M, Kim B H et al. IL-18 induces emphysema and airway and vascular remodeling via IFNγ, IL-17A, and IL-13. Am J Respir Crit Care Med 2012; 1205-1217. Kashmiri S V, De Pascalis R, Gonzales N R, Schlom J. (May 2005). “SDR grafting—a new approach to antibody humanization.”. Methods 36 (1): 25-34 Kawashima, M. et al. Levels of interleukin-18 and its binding inhibitors in the blood circulation of patients with adult-onset Still's disease. Arthritis Rheum 2001; 44(3):550-560. Kim, S. H. et al. Structural requirements of six naturally occurring isoforms of the IL-18 binding protein to inhibit IL-18. Proc Natl Acad Sci USA 2000; 97(3):1190-1195. Kratzer A, Salys J, Nold-Petry El al. Role of IL-18 in second hand smoke-induced emphysema. Am J Respir Cell Mol Biol 2013; 48(6):725-32. Lee C G, Link H, Baluk P, Homer R J, Chapoval S, Bhandari V, Kang M J, Cohn L, Kim Y K, McDonald D M, et al. Vascular endothelial growth factor (VEGF) induces remodeling and enhances TH2-mediated sensitization and inflammation in the lung. Nat Med 2004; 10:1095-1103. Lee C G, Hartl D, Lee G R, Koller B, Matsuura H, Da Silva C A, Sohn M H, Cohn L, Homer R J, Kozhich A A, et al. Role of breast regression protein 39 (BRP-39)/chitinase 3-like-1 in Th2 and IL-13-induced tissue responses and apoptosis. J Exp Med 2009; 206:1149-1166. Liu J et al. Requirement for tumor necrosis factor-receptor 2 in alveolar chemokine expression depends upon the form of the ligand. Am J Respir Cell Mol Biol 2005; 33:463-469. Londhe V A, Maisonet T M, Lopez B, Jeng J M, Li C, Minoo P. A subset of epithelial cells with CCSP promoter activity participates in alveolar development. Am J Respir Cell Mol Biol 2011; 44:804-812. Loza M J, Watt R, Baribaud F, Barnathan E S and Rennard S I. Systemic inflammatory profile and response to anti-tumor necrosis factor therapy in chronic obstructive pulmonary disease. Respir Res 2012; 13:12. Ma B, Kang M J, Lee C G, Chapoval S, Liu W, Chen Q, Coyle A J, Lora J M, Picarella D, Homer R J and Elias J A. Role of CCR5 in IFN-γ-induced and cigarette smoke-induced emphysema. J Clin Invest 2005; 115:3460-3472. Nakajima T and Owen C. Interleukin-18: The Master Regulator Driving Destructive and Remodeling Processes in the Lungs of Patients with Chronic Obstructive Pulmonary Disease? Am J Respir Crit Care Med 2012; 1137-1138. Novick D et al. A novel IL-18BP ELISA shows elevated serum IL-18BP in sepsis and extensive decrease of free IL-18. Cytokine 2001; 14, 334-342. Novick D et al. High circulating levels of free interleukin-18 in patients with active SLE in the presence of elevated levels of interleukin-18 binding protein. J Autoimmun 2010; 34, 121-126. Park M C, Park Y B and Lee S K. Elevated interleukin-18 levels correlated with disease activity in systemic lupus erythematosus. Clin Rheumatol 2004; 23, 225-229. Petersen A M W, Penkowa M and Iversen M. Elevated Levels of IL-18 in Plasma and Skeletal Muscle in Chronic Obstructive Pulmonary Disease. Lung 2007; 161-171. Queen C, Schneider W P, Selick H E, Payne P W, Landolfi N F, Duncan J F, Avdalovic N M, Levitt M, Junghans R P, Waldmann T A. (December 1989). “A humanized antibody that binds to the interleukin 2 receptor.”. Proc Natl Acad Sci USA. 86 (24): 10029-33 Rastrick J M D, Stevenson C S, Eltom S, Grace M, Davies M, et al. Cigarette Smoke Induced Airway Inflammation Is Independent of NF-κB Signalling. PLoS ONE 2013; 8(1):e54128. Ray P, Tang W, Wang P, Homer R, Kuhn C III, Flavell R A and Elias J A. Regulated overexpression of interleukin 11 in the lung: use to dissociate development-dependent and -independent phenotypes. J Clin Invest 1997; 100:2501-2511. Reed L J and Muench H. A simple method of estimating 50% endpoints. Am J Hyg 1938; 27:493-497. Riechmann L, Clark M, Waldmann H, Winter G (1988). “Reshaping human antibodies for therapy”. Nature 332 (6162): 332-323 Rovina N, Dima E and Gerassimou C. Interleukin-18 in induced sputum: Association with lung function in chronic obstructive pulmonary disease. Respiratory Medicine 2009; 1056-1062. Shapiro D S. Transgenic and gene-targeted mice as models for chronic obstructive pulmonary disease. Eur J Respir 2007; 375-378. Taniguchi, M. et al. Characterization of anti-human interleukin-18 (IL-18)/interferon-gamma-inducing factor (IGIF) monoclonal antibodies and their application in the measurement of human IL-18 by ELISA. J Immunol Methods 1997; 206, 107-113. Wang Z, Zheng, Zhu T Z, Homer R J, Riese R J, Chapman H A, Shapiro S D, and Elias J A. Interferon γ induction of pulmonary emphysema in the adult murine lung. J Exp Med 2000; 192:1587-1600. Wright J L, Cosio M and Churg A. Animal models of chronic obstructive disease. Am J Physiol Lung Cell Mol Physiol 2008; L1-L15. Wong C K, Li E K, Ho C Y and Lam C W. Elevation of plasma interleukin-18 concentration is correlated with disease activity in systemic lupus erythematosus. Rheumatology (Oxford) 2000; 39:1078-1081. Zhang J, Dong Z, Zhou R, Luo D, Wei H and Tian Z. Isolation of lymphocytes and their innate immune characterizations from liver, intestine, lung and uterus. Cell Mol Immunol 2005; 2:271-280. Zheng T, Zhu Z, Wang Z, Homer R J, Ma B, Riese R, Chapman H, Shapiro S D and Elias J A. Inducible targeting of IL-13 to the adult lung causes matrix metalloproteinase- and cathepsin-dependent emphysema. J Clin Invest 2000; 106:1081-1093. Zheng T et al. Role of cathepsin S-dependent epithelial cell apoptosis in IFN-gamma-induced alveolar remodeling and pulmonary emphysema. J Immunol 2005; 174:8106-8115.