IL-12/p40 binding proteins
View Patent ↗The present invention encompasses IL-12p40 binding proteins, particularly antibodies that bind human interleukin-12 (hIL-12) and/or human IL-23 (hIL-23). Specifically, the invention relates to antibodies that are chimeric, CDR grafted and humanized antibodies. Preferred antibodies have high affinity for hIL-12 and/or hIL-23 and neutralize h IL-12 and/or hIL-23 activity in vitro and in vivo. An antibody of the invention can be a full-length antibody or an antigen-binding portion thereof. Method of making and method of using the antibodies of the invention are also provided. The antibodies, or antibody portions, of the invention are useful for detecting hIL-12 and/or hIL-23 and for inhibiting hIL-12 and/or hIL-23 activity, e.g., in a human subject suffering from a disorder in which hIL-12 and/or hIL-23 activity is detrimental.
1. An isolated nucleic acid encoding an amino acid sequence of at least one variable domain of a binding protein, wherein said at least one variable domain comprises CDR-H1, CDR-H2, and CDR-H3 amino acid sequences independently selected from the group consisting of:
CDR-H1
CDR-H2
residues 31-37 of SEQ ID NO: 35
residues 52-67 of SEQ ID NO: 35
residues 31-37 of SEQ ID NO: 37
residues 52-67 of SEQ ID NO: 37
residues 31-35 of SEQ ID NO: 39
residues 50-66 of SEQ ID NO: 39
residues 31-35 of SEQ ID NO: 41
residues 50-66 of SEQ ID NO: 41
residues 31-35 of SEQ ID NO: 43
residues 50-66 of SEQ ID NO: 43
residues 31-35 of SEQ ID NO: 45
residues 50-66 of SEQ ID NO: 45
residues 31-35 of SEQ ID NO: 47
residues 50-66 of SEQ ID NO: 47
residues 31-35 of SEQ ID NO: 49
residues 50-66 of SEQ ID NO: 49
residues 31-37 of SEQ ID NO: 51
residues 52-67 of SEQ ID NO: 51
residues 31-35 of SEQ ID NO: 53
residues 47-66 of SEQ ID NO: 53
CDR-H3
residues 100-108 of SEQ ID NO: 35
residues.100-109 of SEQ ID NO: 37
residues 99-106 of SEQ ID NO: 39
residues 99-106 of SEQ ID NO: 41
residues 99-106 of SEQ ID NO: 43
residues 99-101 of SEQ ID NO: 45
residues 99-106 of SEQ ID NO: 47
residues 99-111 of SEQ ID NO: 49
residues 100-109 of SEQ ID NO: 51
residues 99-107 of SEQ ID NO: 53.
2. An isolated nucleic acid encoding an amino acid sequence of at least one variable domain of a binding protein, wherein said at least one variable domain comprises CDR-L1, CDR-L2, and CDR-L3 amino acid sequences independently selected from the group consisting of:
CDR-L1
CDR-L2
CDR-L3
residues 24-34 of SEQ ID NO: 36
residues 50-56 of SEQ ID NO: 36
residues 89-97 of SEQ ID NO: 36
residues 24-34 of SEQ ID NO: 38
residues 50-56 of SEQ ID NO: 38
residues 89-97 of SEQ ID NO: 38
residues 24-34 of SEQ ID NO: 40
residues 50-56 of SEQ ID NO: 40
residues 89-97 of SEQ ID NO: 40
residues 24-34 of SEQ ID NO: 42
residues 50-56 of SEQ ID NO: 42
residues 89-97 of SEQ ID NO: 42
residues 24-34 of SEQ ID NO: 44
residues 50-56 of SEQ ID NO: 44
residues 89-97 of SEQ ID NO: 44
residues 24-34 of SEQ ID NO: 46
residues 50-56 of SEQ ID NO: 46
residues 89-97 of SEQ ID NO: 46
residues 24-34 of SEQ ID NO: 48
residues 50-56 of SEQ ID NO: 48
residues 89-97 of SEQ ID NO: 48
residues 24-38 of SEQ ID NO: 50
residues 53-60 of SEQ ID NO: 50
residues 93-101 of SEQ ID NO: 50
residues 24-34 of SEQ ID NO: 52
residues 50-56 of SEQ ID NO: 52
residues 89-97 of SEQ ID NO: 52
residues 24-34 of SEQ ID NO: 54
residues 50-56 of SEQ ID NO: 54
residues 89-97 of SEQ ID NO: 54.
3. The isolated nucleic acid according to either claim 1 or claim 2 , encoding an antibody construct amino acid sequence, wherein said antibody construct comprises said binding protein and further comprises a linker polypeptide or an immunoglobulin constant domain.
4. The isolated nucleic acid according to either claim 1 or claim 2 encoding an antibody conjugate amino acid sequence, wherein said antibody conjugate comprises said binding protein linked to another moiety.
5. A vector comprising an isolated nucleic acid according to either claim 1 or claim 2 .
6. The vector of claim 5 wherein said vector is selected from the group consisting of pcDNA, pTT, pTT3, pEFBOS, pBV, pJV, and pBJ.
7. A host cell comprising a vector according to claim 5 .
8. The host cell according to claim 7 , wherein said host cell is a prokaryotic cell.
9. The host cell according to claim 8 , wherein said host cell is E. coli.
10. The host cell according to claim 7 , wherein said host cell is a eukaryotic cell.
11. The host cell according to claim 10 , wherein said eukaryotic cell is selected from the group consisting of protist cell, animal cell, plant cell and fungal cell.
12. The host cell according to claim 10 , wherein said eukaryotic cell is an animal cell selected from the group consisting of: a mammalian cell, an avian cell, and an insect cell.
13. The host cell according to claim 10 , wherein said host cell is a CHO cell.
14. The host cell according to claim 10 , wherein said host cell is a COS cell.
15. The host cell according to claim 10 , wherein said host cell is a yeast cell.
16. The host cell according to claim 15 , wherein said yeast cell is Saccharomyces cerevisiae.
17. The host cell according to claim 12 , wherein said host cell is an insect Sf9 cell.
18. A host cell comprising at least one nucleic acid according to either claim 1 or claim 2 .
19. A host cell comprising one or more nucleic acids encoding a polypeptide chain including CDR-H1, CDR-H2, and CDR-H3 amino acid sequences independently selected from the group consisting of:
CDR-H1
CDR-H2
CDR-H3
residues 31-37 of SEQ ID NO: 35
residues 52-67 of SEQ ID NO: 35
residues 100-108 of SEQ ID NO: 35
residues 31-37 of SEQ ID NO: 37
residues 52-67 of SEQ ID NO: 37
residues.100-109 of SEQ ID NO: 37
residues 31-35 of SEQ ID NO: 39
residues 50-66 of SEQ ID NO: 39
residues 99-106 of SEQ ID NO: 39
residues 31-35 of SEQ ID NO: 41
residues 50-66 of SEQ ID NO: 41
residues 99-106 of SEQ ID NO: 41
residues 31-35 of SEQ ID NO: 43
residues 50-66 of SEQ ID NO: 43
residues 99-106 of SEQ ID NO: 43
residues 31-35 of SEQ ID NO: 45
residues 50-66 of SEQ ID NO: 45
residues 99-101 of SEQ ID NO: 45
residues 31-35 of SEQ ID NO: 47
residues 50-66 of SEQ ID NO: 47
residues 99-106 of SEQ ID NO: 47
residues 31-35 of SEQ ID NO: 49
residues 50-66 of SEQ ID NO: 49
residues 99-111 of SEQ ID NO: 49
residues 31-37 of SEQ ID NO: 51
residues 52-67 of SEQ ID NO: 51
residues 100-109 of SEQ ID NO: 51
residues 31-35 of SEQ ID NO: 53
residues 47-66 of SEQ ID NO: 53
residues 99-107 of SEQ ID NO: 53
and nucleic acids encoding a polypeptide chain including CDR-L1, CDR-L2, and CDR-L3 amino acid sequences independently selected from the group consisting of:
CDR-L1
CDR-L2
CDR-L3
residues 24-34 of SEQ ID NO: 36
residues 50-56 of SEQ ID NO: 36
residues 89-97 of SEQ ID NO: 36
residues 24-34 of SEQ ID NO: 38
residues 50-56 of SEQ ID NO: 38
residues 89-97 of SEQ ID NO: 38
residues 24-34 of SEQ ID NO: 40
residues 50-56 of SEQ ID NO: 40
residues 89-97 of SEQ ID NO: 40
residues 24-34 of SEQ ID NO: 42
residues 50-56 of SEQ ID NO: 42
residues 89-97 of SEQ ID NO: 42
residues 24-34 of SEQ ID NO: 44
residues 50-56 of SEQ ID NO: 44
residues 89-97 of SEQ ID NO: 44
residues 24-34 of SEQ ID NO: 46
residues 50-56 of SEQ ID NO: 46
residues 89-97 of SEQ ID NO: 46
residues 24-34 of SEQ ID NO: 48
residues 50-56 of SEQ ID NO: 48
residues 89-97 of SEQ ID NO: 48
residues 24-38 of SEQ ID NO: 50
residues 53-60 of SEQ ID NO: 50
residues 93-101 of SEQ ID NO: 50
residues 24-34 of SEQ ID NO: 52
residues 50-56 of SEQ ID NO: 52
residues 89-97 of SEQ ID NO: 52
residues 24-34 of SEQ ID NO: 54
residues 50-56 of SEQ ID NO: 54
residues 89-97 of SEQ ID NO: 54.
20. A method of producing a protein capable of binding the p40 subunit of IL-12, comprising culturing a host cell of claim 19 in culture medium under conditions sufficient to produce a binding protein capable of binding the p40 subunit of IL-12.
21. A host cell comprising a first nucleic acid encoding a variable heavy chain (VH) set of 3 CDRs and a second nucleic acid encoding a variable light chain (VL) set of 3 CDRs, wherein said VH set of 3 CDRs is selected from any one of the VH sets in the group below and said VL set of 3 CDRs is selected from any one of the VL sets in the group below:
CDR amino acid sequences
VH 1D4 CDR Set
VH 1D4 CDR-H1
residues 31-37 of SEQ ID NO: 35
VH 1D4 CDR-H2
residues 52-67 of SEQ ID NO: 35
VH 1D4 CDR-H3
residues 100-108 of SEQ ID NO: 35
VL 1D4 CDR Set
VL 1D4 CDR-L1
residues 24-34 of SEQ ID NO: 36
VL 1D4 CDR-L2
residues 50-56 of SEQ ID NO: 36
VL 1D4 CDR-L3
residues 89-97 of SEQ ID NO: 36
VH 1A6 CDR Set
VH 1A6 CDR-H1
residues 31-37 of SEQ ID NO: 37
VH 1A6 CDR-H2
residues 52-67 of SEQ ID NO: 37
VH 1A6 CDR-H3
residues 100-109 of SEQ ID NO: 37
VL 1A6 CDR Set
VL 1A6 CDR-L1
residues 24-34 of SEQ ID NO: 38
VL 1A6 CDR-L2
residues 50-56 of SEQ ID NO: 38
VL 1A6 CDR-L3
residues 89-97 of SEQ ID NO: 38
VH 1D8 CDR Set
VH 1D8 CDR-H1
residues 31-35 of SEQ ID NO: 39
VH 1D8 CDR-H2
residues 50-66 of SEQ ID NO: 39
VH 1D8 CDR-H3
residues 99-106 of SEQ ID NO: 39
VL 1D8 CDR Set
VL 1D8 CDR-L1
residues 24-34 of SEQ ID NO: 40
VL 1D8 CDR-L2
residues 50-56 of SEQ ID NO: 40
VL 1D8 CDR-L3
residues 89-97 of SEQ ID NO: 40
VH 3G7 CDR Set
VH 3G7 CDR-H1
residues 31-35 of SEQ ID NO: 41
VH 3G7 CDR-H2
residues 50-66 of SEQ ID NO: 41
VH 3G7 CDR-H3
residues 99-106 of SEQ ID NO: 41
VL 3G7 CDR Set
VL 3G7 CDR-L1
residues 24-34 of SEQ ID NO: 42
VL 3G7 CDR-L2
residues 50-56 of SEQ ID NO: 42
VL 3G7 CDR-L3
residues 89-97 of SEQ ID NO: 42
VH 5E8 CDR Set
VH 5E8 CDR-H1
residues 31-35 of SEQ ID NO: 43
VH 5E8 CDR-H2
residues 50-66 of SEQ ID NO: 43
VH 5E8 CDR-H3
residues 99-106 of SEQ ID NO: 43
VL 5E8 CDR Set
VL 5E8 CDR-L1
residues 24-34 of SEQ ID NO: 44
VL 5E8 CDR-L2
residues 50-56 of SEQ ID NO: 44
VL 5E8 CDR-L3
residues 89-97 of SEQ ID NO: 44
VH 8E1 CDR Set
VH 8E1 CDR-H1
residues 31-35 of SEQ ID NO: 45
VH 8E1 CDR-H2
residues 50-66 of SEQ ID NO: 45
VH 8E1 CDR-H3
residues 99-101 of SEQ ID NO: 45
VL 8E1 CDR Set
VL 8E1 CDR-L1
residues 24-34 of SEQ ID NO: 46
VL 8E1 CDR-L2
residues 50-56 of SEQ ID NO: 46
VL 8E1 CDR-L3
residues 89-97 of SEQ ID NO: 46
VH 1H6 CDR Set
VH 1H6 CDR-H1
residues 31-35 of SEQ ID NO: 47
VH 1H6 CDR-H2
residues 50-66 of SEQ ID NO: 47
VH 1H6 CDR-H3
residues 99-106 of SEQ ID NO: 47
VL 1H6 CDR Set
VL 1H6 CDR-L1
residues 24-34 of SEQ ID NO: 48
VL 1H6 CDR-L2
residues 50-56 of SEQ ID NO: 48
VL 1H6 CDR-L3
residues 89-97 of SEQ ID NO: 48
VH 3A11 CDR Set
VH 3A11 CDR-H1
residues 31-35 of SEQ ID NO: 49
VH 3A11 CDR-H2
residues 50-66 of SEQ ID NO: 49
VH 3A11 CDR-H3
residues 99-111 of SEQ ID NO: 49
VL 3A11 CDR Set
VL 3A11 CDR-L1
residues 24-38 of SEQ ID NO: 50
VL 3A11 CDR-L2
residues 53-60 of SEQ ID NO: 50
VL 3A11 CDR-L3
residues 93-101 of SEQ ID NO: 50
VH 4B4 CDR Set
VH 4B4 CDR-H1
residues 31-37 of SEQ ID NO: 51
VH 4B4 CDR-H2
residues 52-67 of SEQ ID NO: 51
VH 4B4 CDR-H3
residues 100-109 of SEQ ID NO: 51
VL 4B4 CDR Set
VL 4B4 CDR-L1
residues 24-34 of SEQ ID NO: 52
VL 4B4 CDR-L2
residues 50-56 of SEQ ID NO: 52
VL 4B4 CDR-L3
residues 89-97 of SEQ ID NO: 52
VH 7G3 CDR Set
VH 7G3 CDR-H1
residues 31-35 of SEQ ID NO: 53
VH 7G3 CDR-H2
residues 47-66 of SEQ ID NO: 53
VH 7G3 CDR-H3
residues 99-107 of SEQ ID NO: 53
and
VL 7G3 CDR Set
VL 7G3 CDR-L1
residues 24-34 of SEQ ID NO: 54
VL 7G3 CDR-L2
residues 50-56 of SEQ ID NO: 54
VL 7G3 CDR-L3
residues 89-97 of SEQ ID NO: 54.
22. The host cell according to claim 18 , wherein said binding protein comprises at least two variable domain CDR sets.
23. The host cell according to claim 21 , which produces at least two variable domain CDR sets selected from a group consisting of:
VH 1D4 CDR set & VL 1D4 CDR set;
VH 1A6 CDR set & VL 1A6 CDR set;
VH 1D8 CDR set & VL 1D8 CDR set;
VH 3G7 CDR set & VL 3G7 CDR set;
VH 5E8 CDR set & VL 5E8 CDR set;
VH 8E1 CDR set & VL 8E1 CDR set;
VH 1H6 CDR set & VL 1H6 CDR set;
VH 3A11 CDR set & VL 3A11 CDR set;
VH 4B4 CDR set & VL 4B4 CDR set; and
VH 7G3 CDR set & VL 7G3 CDR set.
24. The isolated nucleic acid according to either claim 1 or claim 2 , further encoding a human acceptor framework.
25. The isolated nucleic acid according to claim 24 , wherein said human acceptor framework comprises an amino acid sequence selected from the group consisting of:
SEQ ID NO:6
SEQ ID NO:7
SEQ ID NO:8
SEQ ID NO:9
SEQ ID NO:10
SEQ ID NO:11
SEQ ID NO:12
SEQ ID NO:13
SEQ ID NO:14
SEQ ID NO:15
SEQ ID NO:16
SEQ ID NO:17
SEQ ID NO:18
SEQ ID NO:19
SEQ ID NO:20
SEQ ID NO:21
SEQ ID NO:22
SEQ ID NO:23
SEQ ID NO:24
SEQ ID NO:25
SEQ ID NO:26
SEQ ID NO:27
SEQ ID NO:28
SEQ ID NO:29
SEQ ID NO:30
SEQ ID NO:31
SEQ ID NO:32
SEQ ID NO:33
SEQ ID NO:34
SEQ ID NO:92
SEQ ID NO:93
SEQ ID NO:94
SEQ ID NO:95
SEQ ID NO:96
and
SEQ ID NO:97.
26. The isolated nucleic acid according to claim 24 , wherein said human acceptor framework comprises at least one Framework Region amino acid substitution at a key residue, said key residue selected from the group consisting of:
a residue adjacent to a CDR;
a glycosylation site residue;
a rare residue;
a residue capable of interacting with a p40 subunit of human IL-12;
a residue capable of interacting with a CDR;
a canonical residue;
a contact residue between heavy chain variable region and light chain variable region;
a residue within a Vernier zone; and
a residue in a region that overlaps between a Chothia-defined variable heavy chain CDR1 and a Kabat-defined first heavy chain framework.
27. The isolated nucleic acid according to claim 26 , wherein the key residue is selected from the group consisting of: 3H, 5H, 10H, 11H, 12H, 13H, 15H, 16H, 18H, 19H, 23H, 24H, 25H, 30H, 41H, 44H, 46H, 49H, 66H, 68H, 71H, 73H, 74H, 75H, 76H, 77H, 78H, 79H, 81H, 82H, 82AH, 82BH, 82CH, 83H, 84H, 85H, 86H, 87H, 89H, 93H, 98H, 108H, 109H, 1L, 2L, 3L, 7L, 8L, 9L, 10L, 11L, 12L, 13L, 15L, 17L, 19L, 20L, 21L, 22L, 36L, 41L, 42L, 43L, 45L, 46L, 58L, 60L, 62L, 63L, 67L, 70L, 73L, 74L, 77L, 78L, 79L, 80L, 83L, 85L, 87L, 104L, and 106L.
28. An isolated nucleic acid encoding an amino acid sequence of at least one variable domain of a binding protein, wherein said at least one variable domain comprises an amino acid sequence selected from the group consisting of:
SEQ ID NO:61
SEQ ID NO:62
SEQ ID NO:63
SEQ ID NO:64
SEQ ID NO:65
SEQ ID NO:66
SEQ ID NO:67
SEQ ID NO:68
SEQ ID NO:69
SEQ ID NO:70
SEQ ID NO:71
SEQ ID NO:72
SEQ ID NO:73
SEQ ID NO:74
SEQ ID NO:75
SEQ ID NO:76
SEQ ID NO:77
and
SEQ ID NO:78.
29. The host cell according to claim 18 , producing a binding protein comprising two variable domains, wherein said two variable domains comprise amino acid sequences selected from the group consisting of:
SEQ ID NO:61 & SEQ ID NO:62,
SEQ ID NO:63 & SEQ ID NO:64,
SEQ ID NO:65 & SEQ ID NO:66,
SEQ ID NO:67 & SEQ ID NO:68,
SEQ ID NO:69 & SEQ ID NO:70,
SEQ ID NO:71 & SEQ ID NO:72,
SEQ ID NO:73 & SEQ ID NO:74,
SEQ ID NO:75 & SEQ ID NO:76,
SEQ ID NO:77 & SEQ ID NO:78
SEQ ID NO:67 & SEQ ID NO:70, and
SEQ ID NO:69 & SEQ ID NO:68.
30. The host cell according to claim 18 , producing a binding protein comprising two variable domains, wherein said two variable domains comprise amino acid sequences selected from the group consisting of:
SEQ ID NO:67 & SEQ ID NO:79,
SEQ ID NO:80 & SEQ ID NO:81,
SEQ ID NO:82 & SEQ ID NO:83,
SEQ ID NO:84 & SEQ ID NO:85,
SEQ ID NO:86 & SEQ ID NO:87,
SEQ ID NO:88 & SEQ ID NO:89,
SEQ ID NO:90 & SEQ ID NO:91,
SEQ ID NO:98 & SEQ ID NO:99,
SEQ ID NO:100 & SEQ ID NO:101,
SEQ ID NO:102 & SEQ ID NO:103,
SEQ ID NO:104 & SEQ ID NO:105,
SEQ ID NO:106 & SEQ ID NO:107, and
SEQ ID NO:108 & SEQ ID NO:109.
31. The host cell according to claim 29 , wherein said binding protein is capable of binding a target selected from the group consisting of IL-12, IL-23, or both.
32. The host cell according to claim 30 , wherein said binding protein is capable of binding a target selected from the group consisting of IL-12, IL-23, or both.
33. The host cell according to claim 32 , wherein said two variable domains comprise: SEQ ID NO:82 & SEQ ID NO:83.