Polynucleotides encoding humanized anti-factor D antibodies
The invention relates to anti-Factor D antibodies, their nucleic acid and amino acid sequences, the cells and vectors that harbor these antibodies and their production and their use in the preparation of compositions and medicaments for treatment of diseases and disorders associated with excessive or uncontrolled complement activation. These antibodies are useful for diagnostics, prophylaxis and treatment of disease.
1. An isolated polynucleotide encoding an anti-Factor D antibody or an antigen-binding fragment thereof, wherein the anti-Factor D antibody or an antigen-binding fragment thereof comprises:
(a) a light chain variable domain comprising: (1) light chain HVR-1 (HVR-L1) comprising the sequence ITSTDIDDDMN (SEQ ID NO: 30) or a sequence that differs from SEQ ID NO: 30 by one or more amino acid substitutions, wherein said one or more substitutions are substitution of the amino acid residue at position 10 of SEQ ID NO: 30 to L or I and/or substitution of the amino acid at position 11 of SEQ ID NO: 30 to A or Q, (2) light chain HVR-2 (HVR-L2) comprising the sequence GGNTLRP (SEQ ID NO: 35) or a sequence that differs from SEQ ID NO: 35 by one amino acid substitution, wherein said one substitution is substitution of the amino acid residue at position 3 of SEQ ID NO: 35 to S or A, and, (3) light chain HVR-3 (HVR-L3) comprising the sequence LQSDSLPYT (SEQ ID NO: 38); and
(b) a heavy chain variable domain comprising: (1) heavy chain HVR-1 (HVR-H1) comprising the sequence GYTFTNYGMN (SEQ ID NO: 39), (2) heavy chain HVR-2 (HVR-H2) comprising the sequence WINTYTGETTYADDFKG (SEQ ID NO: 40), and (3) heavy chain HVR-3 (HVR-H3) comprising the sequence EGGVNN (SEQ ID NO: 41) or a sequence that differs from SEQ ID NO: 41 by one or more amino acid substitutions, wherein said one or more substitutions are substitution of the amino acid residue at position 5 of SEQ ID NO: 41 with A or Q and/or substitution of the amino acid residue at position 6 of SEQ ID NO: 41 with A or Q,
wherein said antibody or antigen-binding fragment thereof comprises at least one of the substitutions set forth in (a) (1), (a) (2), or (b) (3) above and/or the heavy chain variable domain comprises an E at position 1.
2. The polynucleotide of claim 1 , wherein the anti-Factor D antibody or antigen-binding fragment thereof comprises an E at position 1 of the heavy chain variable domain.
3. The polynucleotide of claim 2 , wherein the light chain variable domain comprises the amino acid sequence of
DIQVTQSPSSLSASVGDRVTITCITSTDIDDX 4 X 5 WYQQKPGKVPKLLISGGX 6 -TLRPGVP SRFSGSGSGTDFTLTISSLQPEDVATYYCLQSDSLPYTFGQGTKX 7 EIK (SEQ ID NO: 73), wherein X 4 is M, L or I; X 5 is N, A or Q; X 6 is N, S or A; and X 7 is L or V.
4. The polynucleotide of claim 3 , wherein X 7 is V.
5. The polynucleotide of claim 1 , wherein (a) the heavy chain variable domain comprises: i. an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 39; ii. an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 40; iii. an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44 or SEQ ID NO: 45; and (b) the light chain variable domain comprises: iv. an HVR-L1 comprising the amino acid sequence selected from SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33 and SEQ ID NO: 34; v. an HVR-L2 comprising the amino acid sequence of SEQ ID NO: 35, SEQ ID NO: 36 or SEQ ID NO: 37; and vi. an HVR-L3 comprising the amino acid sequence of SEQ ID NO: 38.
6. The polynucleotide of claim 5 , wherein the HVR-H3 comprises the amino acid sequence of SEQ ID NO: 41.
7. The polynucleotide of claim 5 , wherein the HVR-L1 comprises the amino acid sequence of SEQ ID NO: 30.
8. The polynucleotide of claim 5 , wherein the HVR-L2 comprises the amino acid sequence of SEQ ID NO: 35.
9. The polynucleotide of claim 5 , wherein the HVR-L1 comprises the amino acid sequence of SEQ ID NO: 30, the HVR-L2 comprises the amino acid sequence of SEQ ID NO: 35, and the HVR-L3 comprises the amino acid sequence of SEQ ID NO: 38.
10. The polynucleotide of claim 5 , wherein the HVR-H1 comprises the amino acid sequence of SEQ ID NO: 39, the HVR-H2 comprises the amino acid sequence of SEQ ID NO: 40, the HVR-13 comprises the amino acid sequence of SEQ ID NO: 41, the HVR-L1 comprises the amino acid sequence of SEQ ID NO: 30, the HVR-L2 comprises the amino acid sequence of SEQ ID NO: 35, and the HVR-L3 comprises the amino acid sequence of SEQ ID NO: 38.
11. The polynucleotide of claim 10 , wherein the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 1, and wherein the amino acid residue at position 104 of SEQ ID NO: 1 is substituted with V.
12. The polynucleotide of claim 11 , wherein the amino acid residue at position 1 of the heavy chain variable domain is an E.
13. The polynucleotide of claim 1 , wherein the heavy chain variable domain comprises the sequence of SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28 or SEQ ID NO: 29.
14. The polynucleotide of claim 1 , wherein the light chain variable domain comprises the sequence of 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, or SEQ ID NO: 14.
15. An isolated polynucleotide encoding an anti-Factor D antibody or an antigen-binding fragment thereof, wherein the anti-Factor D antibody or the antigen-binding fragment thereof comprises a polypeptide comprising the following amino acid sequence:
X 1 VQLVQSGPELKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTY TGETTYADDFKGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCEREGGVX 2 X 3 WGQGTV TVSS (SEQ ID NO: 74), wherein X 1 is E; X 2 is N, A or Q; and X 3 is N, A or Q.
16. The polynucleotide of claim 15 , wherein the anti-Factor D antibody or antigen-binding fragment thereof further comprises a polypeptide comprising the following amino acid sequence:
DIQVTQSPSSLSASVGDRVTITCITSTDIDDDX 4 X 5 WYQQKPGKVPKLLISGG-X 6 TLRPGVP SRFSGSGSGTDFTLTISSLQPEDVATYYCLQSDSLPYTFGQGTKX 7 EIK (SEQ ID NO:73), wherein X 4 is M, L or I; X 5 is N, A or Q; X 6 is N, S or A; and X 7 is L or V.
17. An isolated polynucleotide encoding an anti-Factor D antibody or an antigen-binding fragment thereof, wherein the anti-Factor I) antibody or an antigen-binding fragment thereof comprises the heavy chain variable domain sequence of SEQ ID NO: 19.
18. The polynucleotide of claim 17 , wherein the anti-Factor D antibody or antibody fragment thereof further comprises the light chain variable domain sequence of SEQ ID NO: 7.
19. An isolated polynucleotide encoding an anti-Factor D antibody or an antigen-binding fragment thereof, wherein the anti-Factor D antibody or an antigen-binding fragment thereof comprises the heavy chain sequence of SEQ ID NO: 63.
20. The polynucleotide of claim 19 , wherein the anti-Factor D antibody further comprises the light chain sequence of SEQ ID NO: 61.
21. An isolated polynucleotide encoding an anti-Factor D antibody or an antigen-binding fragment thereof, wherein the anti-Factor D antibody or an antigen-binding fragment thereof comprises: (a) a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 1, wherein the amino acid residue at position 104 of SEQ ID NO: 1 is substituted with V; and (b) a heavy chain variable domain comprising an HVR-H1 comprising the amino acid sequence of SEQ ID NO: 39, an HVR-H2 comprising the amino acid sequence of SEQ ID NO: 40, and an HVR-H3 comprising the amino acid sequence of SEQ ID NO: 41, wherein the amino acid residue at position 1 of the heavy chain variable domain is an E.
22. A vector comprising the polynucleotide of claim 1 , 15 , 17 , 19 , or 21 .
23. An isolated host cell comprising the vector of claim 22 .
24. The host cell of claim 23 , wherein the host cell is eukaryotic.
25. The host cell of claim 24 , wherein the host cell is a CHO cell.
26. The host cell of claim 23 , wherein the host cell is prokaryotic.
27. A method of making an anti-Factor D antibody, wherein the method a comprises (a) culturing the host cell of claim 23 under conditions suitable for expression of the polynucleotide encoding the antibody, and (b) isolating the antibody.