Anti-transthyretin antibodies
The invention provides antibodies that specifically bind transthyretin (TTR). The antibodies can be used for treating or effecting prophylaxis of diseases or disorders associated with TTR accumulation or accumulation of TTR deposits (e.g., TTR amyloidosis). The antibodies can also be used for diagnosing TTR amyloidosis and inhibiting or reducing aggregation of TTR, among other applications.
1. A vector comprising a nucleic acid encoding a mature heavy chain variable region and a mature light chain variable region operably linked to one or more regulatory sequences to effect expression in a mammalian cell of an antibody comprising the mature heavy chain variable region and mature light chain variable region that binds to human transthyretin, wherein the mature heavy chain variable region comprises three heavy chain CDRs having sequences comprising SEQ ID NOS:67-69, respectively, and the mature light chain variable region comprises three light chain CDRs having sequences comprising SEQ ID NOS:77-79, respectively, except that positions H52 and L26 by Kabat numbering are independently each N or S.
2. The vector of claim 1 wherein the mature heavy chain variable region is encoded by a sequence comprising any one of SEQ ID NOS:44-51 and SEQ ID NOS:91-93 and the nucleic acid encoding the mature light chain variable region is encoded by a sequence comprising any one of SEQ ID NOS:52-56 and SEQ ID NOS:94-96.
3. The vector of claim 1 , wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOS:5-12 and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOS:19-23.
4. The vector of claim 3 , wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:11 and the mature light chain variable region has the amino acid sequence of SEQ ID NO:19.
5. The vector of claim 1 , wherein positions H47 and H108 in the mature heavy chain variable region are each occupied by L, and wherein position L36 in the mature light chain variable region is occupied by F, wherein positions are defined by Kabat numbering.
6. The vector of claim 2 having a sequence comprising SEQ ID NO:50 encoding the mature heavy chain variable region and SEQ ID NO:52 encoding the mature light chain variable region.
7. The vector of claim 1 , wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOS:64-66, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOS:74-76.
8. The vector of claim 7 , wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:65 and the mature light chain variable region has the amino acid sequence of SEQ ID NO:76.
9. The vector of claim 1 , wherein positions H1 and H47 in the mature heavy chain variable region are occupied by E and L, respectively, and wherein positions L36 and L60 in the mature light chain variable region are occupied by F and S, respectively, wherein positions are defined by Kabat numbering.
10. The vector of claim 2 having a sequence comprising SEQ ID NO:92 encoding the mature heavy chain variable region and SEQ ID NO:96 encoding the mature light chain variable region.
11. The vector of claim 1 , wherein the nucleic acid further encodes a heavy chain constant region fused to the mature heavy chain variable region and a light chain constant region fused to the mature light chain variable region.
12. The vector of claim 11 , wherein the heavy chain constant region has the sequence of any one of SEQ ID NOS:101-103 with or without the C-terminal lysine and the light chain constant region has the sequence of any one of SEQ ID NOS:104-105.
13. The vector of claim 11 , wherein the heavy chain constant region is encoded by the sequence of SEQ ID NO:106 and the light chain constant region is encoded by the sequence of any one of SEQ ID NOS:107-108.
14. The vector of claim 1 , wherein the antibody is a scFv.
15. The vector of claim 1 , wherein the antibody is a Fab fragment.
16. The vector of claim 1 , wherein the one or more regulatory sequences include one or more of a promoter, enhancer, ribosome binding site, and transcription termination signal.
17. The vector of claim 1 , wherein the nucleic acid further encodes signal peptides fused to the mature heavy and light chain variable regions.
18. The vector of claim 1 , wherein the nucleic acid is codon-optimized for expression in a host cell.
19. The vector of claim 1 , wherein the one or more regulatory sequences include a eukaryotic promoter.
20. The vector of claim 1 , wherein the nucleic acid further encodes a selection marker.
21. A host cell comprising the nucleic acid of claim 1 .
22. A method of expressing an antibody in a mammalian cell comprising incorporating the nucleic acid of claim 1 into the genome of a transgenic animal, whereby the antibody is expressed.
23. A host cell comprising first and second nucleic acids respectively encoding a mature heavy chain variable region and a mature light chain variable region, each operably linked to one or more regulatory sequences to effect expression of an antibody comprising the mature heavy chain variable region and mature light chain variable region, wherein the mature heavy chain variable region comprises three heavy chain CDRs having sequences comprising SEQ ID NOS:67-69, respectively, and the mature light chain variable region comprises three light chain CDRs having sequences comprising SEQ ID NOS:77-79, respectively, except that positions H52 and L26 by Kabat numbering are independently each N or S.
24. The host cell of claim 23 , wherein the mature heavy chain variable region is encoded by a sequence comprising any one of SEQ ID NOS:44-51 and SEQ ID NOS:91-93 and the nucleic acid encoding the mature light chain variable region is encoded by a sequence comprising any one of SEQ ID NOS:52-56 and SEQ ID NOS:94-96.
25. The host cell of claim 23 , wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOS:5-12 and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOS:19-23.
26. The host cell of claim 25 , wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:11 and the mature light chain variable region has the amino acid sequence of SEQ ID NO:19.
27. The host cell of claim 23 , wherein positions H47 and H108 in the mature heavy chain variable region are each occupied by L, and wherein position L36 in the mature light chain variable region is occupied by F, wherein positions are defined by Kabat numbering.
28. The host cell of claim 24 , wherein the nucleic acid encoding the mature heavy chain variable region has a sequence comprising SEQ ID NO:50 and the nucleic acid encoding the mature light chain variable region has a sequence comprising SEQ ID NO:52.
29. The host cell of claim 23 , wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOS:64-66, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOS:74-76.
30. The host cell of claim 29 , wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:65 and the mature light chain variable region has the amino acid sequence of SEQ ID NO:76.
31. The host cell of claim 23 , wherein positions H1 and H47 in the mature heavy chain variable region are occupied by E and L, respectively, and wherein positions L36 and L60 in the mature light chain variable region are occupied by F and S, respectively, wherein positions are defined by Kabat numbering.
32. The host cell of claim 24 , wherein the nucleic acid encoding the mature heavy chain variable region has a sequence comprising SEQ ID NO:92 and the nucleic acid encoding the mature light chain variable region has a sequence comprising SEQ ID NO:96.
33. The host cell of claim 23 , wherein the nucleic acids respectively further encode a heavy chain constant region fused to the mature heavy chain variable region and a light chain constant region fused to the mature light chain variable region.
34. The host cell of claim 33 , wherein the heavy chain constant region has the sequence of any one of SEQ ID NOS:101-103 with or without the C-terminal lysine and the light chain constant region has the sequence of any one of SEQ ID NOS:104-105.
35. The host cell of claim 33 , wherein the heavy chain constant region is encoded by the sequence of SEQ ID NO:106 and the light chain constant region is encoded by the sequence of any one of SEQ ID NOS:107-108.
36. A method of expressing an antibody in a mammalian cell comprising incorporating first and second nucleic acids respectively encoding a mature heavy chain variable region and a mature light chain variable region, each operably linked to one or more regulatory sequences to effect expression of an antibody comprising the mature heavy chain variable region and mature light chain variable region, wherein the mature heavy chain variable region comprises three heavy chain CDRs having sequences comprising SEQ ID NOS:67-69, respectively, and the mature light chain variable region comprises three light chain CDRs having sequences comprising SEQ ID NOS:77-79, respectively, except that positions H52 and L26 by Kabat numbering are independently each N or S into the genome of a transgenic animal, whereby the antibody is expressed.