Antagonistic anti-human CD40 monoclonal antibody
The present invention provides novel antagonistic anti-human CD40 monoclonal antibodies, methods for generating them and uses thereof.
1. A polypeptide comprising an amino acid sequence
1 11 21 31
| | | |
GFSX 1 S RYSVY WX 2 RQP PGKGX 3 EWX 4 GM MWGGG STDYS
41 51 61
| | |
TSLKS RLTIS KDTSK SQVX 5 L KMNSL RTDDT AMYYC
71
|
VRTDG DY
wherein:
X 1 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H;
X 2 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H;
X 3 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H;
X 4 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H; and
X 5 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H
(SEQ ID NO. 1).
2. A polypeptide according to claim 1 , wherein:
X 1 is G, A, V, L, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H.
3. A polypeptide according to claim 1 , wherein:
X 1 is G, A, V, L, P, F or M;
X 2 is G, A, V, L, I, P, F or M;
X 3 is G, A, V, L, I, P, F, M;
X 4 is G, A, V, L, I, P, F, M; and
X 5 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T or Y.
4. A polypeptide according to claim 1 , wherein:
X 1 is G, A, V, L or M;
X 2 is G, A, V, L, I or M;
X 3 is G, +A, V, L, I, P, F, M;
X 4 is G, A, V, L, I or M; and
X 5 is P, F, W, N, Q, S, T or Y.
5. A polypeptide according to claim 4 , wherein:
X 1 is L;
X 2 is I;
X 3 is P;
X 4 is M; or X 5 is S.
6. A polypeptide according to claim 1 , wherein:
X 1 is I and X 2 is V;
X 1 is L and X 2 is I;
X 1 is V and X 2 is V;
X 1 is L and X 2 is L; or
X 1 is L and X 2 is V.
7. A polypeptide according to claim 6 , wherein
X 3 is P;
X 4 is M; and
X 5 is F or X 5 is S, preferably S.
8. A polypeptide according to claim 1 , wherein:
X 1 is L; X 2 is V; X 3 is L; X 4 is L and X 5 is F.
9. A polypeptide comprising amino acid sequence
1 11
| |
GFSX 1 S RYSVY WX 2 R
wherein:
X 1 is L and X 2 is I (SEQ ID NO: 3); or
X 1 is I and X 2 is V (SEQ ID NO: 4).
10. An anti-CD40 antibody comprising a polypeptide according to claim 1 or claim 8 .
11. An anti-CD40 antibody according to claim 10 , comprising a constant region of a human antibody, preferably an IgG constant region.
12. An anti-CD40 antibody according to claim 10 , wherein said constant region is a region that is deficient in complement activation, preferably human IgG 4 constant region or a mutated human IgG 1 constant region.
13. An antagonistic anti-human CD40 monoclonal antibody comprising a polypeptide according to claim 1 or claim 8 .
14. An antagonistic anti-human CD40 monoclonal antibody according to claim 13 , wherein the antibody is deimmunized.
15. An antagonistic anti-human CD40 monoclonal antibody according to claim 13 comprising an amino acid sequence
1 11 21 31
| | | |
X 6 LGX 7 X 8 ASISC RSSQS LX 9 NSN GNTYL HWYLQ RPGQS
41 51 61 71
| | | |
PRLLI YKVSN RFSGV PDRFS GSGSG TDFTL KISRV EAEDX 10
81 91
| |
GVYX 11 C SQSTH VPWT
wherein:
X 6 is N, Q, S, T, Y, W or C;
X 7 is D, E, N, Q, S, T, Y, W or C;
X 8 is N, Q, S, T, Y, G, A, V, L, I, P, F, M, W or C;
X 9 is G, A, V, L, I, P, F, M;
X 10 is G, A, V, L, I, P, F, M; and
X 11 is N, Q, S, T, Y, G, A, V, L, I, P, F, M, W or C
(SEQ ID NO: 5).
16. An antagonistic anti-human CD40 monoclonal antibody according to claim 15 comprising an amino acid sequence
1 11 21 31
| | | |
ELQLT QSPLS LPVX 6 L GX 7 X 8 AS ISCRS SQSLX 9 NSNGN
41 51 61 71
| | | |
TYLHW YLQRP GQSPR LLIYK VSNRF SGVPD RFSGS GSGTD
81 91 101 111
| | | |
FTLKI SRVEA EDX 10 GV YX 11 CSQ STHVP WTFGG GTKLE IKR
(SEQ ID NO: 6).
17. An antibody according to claim 15 or claim 16 wherein
X 6 is T or S, X 7 is D or Q, X 8 is Q or P, X 9 is V or A, X 10 is V or L and X 11 is F or Y.
18. An antibody according to claim 15 or claim 16 , wherein:
X 6 is T, X 7 is Q, X 8 is P, X 9 is A, X 10 is V and X 1 , is Y.
19. An antibody obtainable by a method for producing an antibody comprising culturing a cell comprising an antibody according to claim 15 or claim 16 and harvesting said antibody from said culture.
20. An antibody according to claim 19 that is purified.
21. A polypeptide comprising an amino acid sequence
1 11 21 31
| | | |
QVKLQ ESGPG LVKPS ETLSI TCTVS GFSX 1 S RYSVY WX 2 RQP
41 51 61 71
| | | |
PGKGP EWX 4 GM MWGGG STDYS TSLKS RLTIS KDTSK
81 91 101 111
| | | |
SQVX 5 L KMNSL RTDDT AMYYC VRTDG DYWGQ GTTVT VSS.
wherein:
X 1 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H;
X 2 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H;
X 3 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H;
X 4 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H; and
X 5 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H
(SEQ ID NO. 2).
22. A polypeptide according to claim 21 , wherein:
i. X 1 is G, A, V, L, P, F, M, W, C, N, Q, S, T, Y, D, E, K, R or H.
23. A polypeptide according to claim 21 , wherein:
i. X 1 is G, A, V, L, P, F or M;
ii. X 2 is G, A, V, L, I, P, F or M;
iii. X 3 is G, A, V, L, I, P, F, M;
iv. X 4 is G, A, V, L, I, P, F, M; and
v. X 5 is G, A, V, L, I, P, F, M, W, C, N, Q, S, T or Y.
24. A polypeptide according to claim 21 , wherein:
i. X 1 is G, A, V, L or M;
ii. X 2 is G, A, V, L, I or M;
iii. X 3 is G, A, V, L, I, P, F, M;
iv. X 4 is G, A, V, L, I or M; and
v. X 5 is P, F, W, N, Q, S, T or Y.
25. A polypeptide according to claim 24 , wherein:
i. X 1 is L;
ii. X 2 is I;
iii. X 3 is P;
iv. X 4 is M; or X 5 is S.
26. A polypeptide according to claim 21 , wherein:
i. X 1 is I and X 2 is V;
ii. X 1 is L and X 2 is I;
iii. X 1 is V and X 2 is V;
iv. X 1 is L and X 2 is L; or
v. X 1 is L and X 2 is V.
27. A polypeptide according to claim 26 , wherein
i. X 3 is P;
ii. X 4 is M; and
iii. X 5 is F or X 5 is S, preferably S.
28. A polypeptide according to claim 21 , wherein:
i. X 1 is L; X 2 is V; X 3 is L; X 4 is L and X 5 is F.
29. An anti-CD40 antibody comprising a polypeptide according to claim 21 .
30. An anti-CD40 antibody according to claim 29 , comprising a constant region of a human antibody, preferably an IgG constant region.
31. An anti-CD40 antibody according to claim 29 , wherein said constant region is a region that is deficient in complement activation, preferably human IgG 4 constant region or a mutated human IgG 1 constant region.
32. An antagonistic anti-human CD40 monoclonal antibody comprising a polypeptide according to claim 21 .
33. An antagonistic anti-human CD40 monoclonal antibody according to claim 32 further comprising an amino acid sequence
1 11 21 31
| | | |
X 6 LGX 7 X 8 ASISC RSSQS LX 9 NSN GNTYL HWYLQ RPGQS
41 51 61 71
| | | |
PRLLI YKVSN RFSGV PDRFS GSGSG TDFTL KISRV EAEDX 10
81 91
| |
GVYX 11 C SQSTH VPWT
wherein:
X 6 is N, Q, S, T, Y, W or C;
X 7 is D, E, N, Q, S, T, Y, W or C;
X 8 is N, Q, S, T, Y, G, A, V, L, I, P, F, M, W or C;
X 9 is G, A, V, L, I, P, F, M;
X 10 is G, A, V, L, I, P, F, M; and
X 11 is N, Q, S, T, Y, G, A, V, L, I, P, F, M, W or C
(SEQ ID NO: 5).
34. An antagonistic anti-human CD40 monoclonal antibody according to claim 32 further comprising an amino acid sequence
1 11 21 31
| | | |
ELQLT QSPLS LPVX 6 L GX 7 X 8 AS ISCRS SQSLX 9 NSNGN
41 51 61 71
| | | |
TYLHW YLQRP GQSPR LLIYK VSNRF SGVPD RFSGS GSGTD
81 91 101
| | | |
FTLKI SRVEA EDX 10 GV YX 11 CSQ STHVP WTFGG GTKLE
111
IKR (SEQ ID NO: 6).
35. An antagonistic anti-human CD40 monoclonal antibody according to claim 33 or claim 34 wherein
X 6 is T or S, X 7 is D or Q, X 8 is Q or P, X 9 is V or A, X 10 is V or L and X 11 is F or Y.
36. An antagonistic anti-human CD40 monoclonal antibody according to claim 35 , wherein:
X 6 is T, X 7 is Q, X 8 is P, X 9 is A, X 10 is V and X 11 is Y.
37. An antibody obtainable by culturing a cell comprising an antibody according to claim 33 or claim 34 and harvesting said antibody from said culture.
38. An antibody according to claim 37 , wherein the antibody is purified.