Human anti-alpha-synuclein antibodies
Provided are human alpha-synuclein-specific autoantibodies as well as fragments, derivatives and variants thereof as well as methods related thereto. Assays, kits, and solid supports related to antibodies specific for α-synuclein are also disclosed. The antibody, immunoglobulin chain(s), as well as binding fragments, derivatives and variants thereof can be used in pharmaceutical and diagnostic compositions for α-synuclein targeted immunotherapy and diagnosis, respectively.
1. An expression vector comprising a promoter operably linked to a polynucleotide encoding a polypeptide comprising:
an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively, wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO:13 binds to human alpha-synuclein;
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:10 binds to human alpha-synuclein;
an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:3, residues 50-66 of SEQ ID NO:3, and residues 99-110 of SEQ ID NO:3, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:7 binds to human alpha-synuclein;
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:6, residues 49-55 of SEQ ID NO:6, and residues 88-98 of SEQ ID NO:6, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:4 binds to human alpha-synuclein;
an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:15, residues 50-66 of SEQ ID NO:15, and residues 99-112 of SEQ ID NO:15, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:19 binds to human alpha-synuclein;
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 24-34 of SEQ ID NO:18, residues 50-56 of SEQ ID NO:18, and residues 89-96 of SEQ ID NO:18, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:16 binds to human alpha-synuclein; or
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 24-34 of SEQ ID NO:21, residues 50-56 of SEQ ID NO:21, and residues 89-97 of SEQ ID NO:21, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:16 binds to human alpha-synuclein.
2. The expression vector of claim 1 , wherein the promoter is a heterologous promoter.
3. The expression vector of claim 2 , wherein the heterologous promoter is a cytomegalovirus, simian virus 40, or retroviral promoter.
4. The expression vector of claim 2 , wherein the heterologous promoter is the cytomegalovirus immediate early promoter.
5. The expression vector of claim 4 , wherein the expression vector comprises cytomegalovirus intron-A.
6. The expression vector of claim 1 , wherein the polypeptide comprises a signal peptide.
7. The expression vector of claim 6 , wherein the signal peptide is a heterologous signal peptide.
8. The expression vector of claim 1 , wherein the polypeptide comprises:
an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO:10;
an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO:13;
an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO:4;
an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO:7;
an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO:16;
an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO:19; or
an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO:22.
9. The expression vector of claim 1 , wherein the polynucleotide comprises the nucleic acid sequence set forth in:
SEQ ID NO:8; or
SEQ ID NO:11.
10. The expression vector of claim 1 , wherein the expression vector is a plasmid, phage, virus, or retrovirus.
11. An expression vector comprising:
a first polynucleotide encoding a first polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively; and
a second polynucleotide encoding a second polypeptide comprising an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively,
wherein the immunoglobulin heavy chain or fragment thereof when paired with the immunoglobulin light chain or fragment thereof forms an anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof.
12. The expression vector of claim 11 , wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.
13. The expression vector of claim 11 , wherein:
the VH consists of the amino acid sequence set forth in SEQ ID NO:10; and
the VL consists of the amino acid sequence set forth in SEQ ID NO:13.
14. The expression vector of claim 11 , wherein:
the first polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO:8; and
the second polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO:11.
15. The expression vector of claim 11 , wherein the expression vector is a plasmid, phage, virus, or retrovirus.
16. A cDNA comprising a polynucleotide encoding a polypeptide comprising:
an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively, wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO:13 binds to human alpha-synuclein;
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:10 binds to human alpha-synuclein;
an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:3, residues 50-66 of SEQ ID NO:3, and residues 99-110 of SEQ ID NO:3, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:7 binds to human alpha-synuclein;
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:6, residues 49-55 of SEQ ID NO:6, and residues 88-98 of SEQ ID NO:6, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:4 binds to human alpha-synuclein;
an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:15, residues 50-66 of SEQ ID NO:15, and residues 99-112 of SEQ ID NO:15, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:19 binds to human alpha-synuclein;
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 24-34 of SEQ ID NO:18, residues 50-56 of SEQ ID NO:18, and residues 89-96 of SEQ ID NO:18, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:16 binds to human alpha-synuclein; or
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 24-34 of SEQ ID NO:21, residues 50-56 of SEQ ID NO:21, and residues 89-97 of SEQ ID NO:21, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:16 binds to human alpha-synuclein.
17. A host cell comprising:
a first expression vector comprising a first polynucleotide encoding a first polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively; and
a second expression vector comprising a second polynucleotide encoding a second polypeptide comprising an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively,
wherein the immunoglobulin heavy chain or fragment thereof when paired with the immunoglobulin light chain or fragment thereof forms an anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof.
18. The host cell of claim 17 , wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.
19. The host cell of claim 17 , wherein the host cell is a mammalian host cell.
20. The host cell of claim 17 , wherein the host cell is a Chinese Hamster Ovary (CHO) cell, a HEK 293 cell, or a NSO cell.
21. A host cell comprising an expression vector comprising:
a first polynucleotide encoding a first polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively; and
a second polynucleotide encoding a second polypeptide comprising an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively,
wherein the immunoglobulin heavy chain or fragment thereof when paired with the immunoglobulin light chain or fragment thereof forms an anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof.
22. The host cell of claim 21 , wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.
23. The host cell of claim 21 , wherein the host cell is a mammalian host cell.
24. The host cell of claim 21 , wherein the host cell is a Chinese Hamster Ovary (CHO) cell, a HEK 293 cell, or a NSO cell.
25. A method for preparing an anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof, the method comprising:
culturing the host cell of claim 17 in a cell culture; and
isolating the anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof from the cell culture.
26. The method of claim 25 , further comprising formulating the anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof into a sterile pharmaceutical composition suitable for administration to a human subject.
27. The method of claim 26 , wherein the pharmaceutical composition is suitable for intravenous or subcutaneous administration.
28. A method for preparing an anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof, the method comprising:
culturing the host cell of claim 21 in a cell culture; and
isolating the anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof from the cell culture.
29. The method of claim 28 , further comprising formulating the anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof into a sterile pharmaceutical composition suitable for administration to a human subject.
30. The method of claim 29 , wherein the pharmaceutical composition is suitable for intravenous or subcutaneous administration.
31. A method of preparing a pharmaceutical composition, the method comprising formulating an anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof into a sterile pharmaceutical composition suitable for administration to a human subject, wherein the anti-human alpha-synuclein antibody or human alpha-synuclein-binding fragment thereof comprises (i) an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively; and (ii) an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively.
32. The method of claim 31 , wherein (i) the VH consists of the amino acid sequence set forth in SEQ ID NO:10; and (ii) the VL consists of the amino acid sequence set forth in SEQ ID NO:13.
33. The method of claim 32 , wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.
34. The method of claim 33 , wherein the pharmaceutical composition is suitable for intravenous or subcutaneous administration.
35. The expression vector of claim 1 , wherein the polypeptide comprises:
an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:13 binds to human alpha-synuclein.
36. The expression vector of claim 1 , wherein the polypeptide comprises:
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:10 binds to human alpha-synuclein.
37. The expression vector of claim 1 , wherein the polypeptide comprises:
an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO:10.
38. The expression vector of claim 1 , wherein the polypeptide comprises:
an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO:13.
39. The expression vector of claim 10 , wherein the expression vector is a plasmid.
40. The expression vector of claim 15 , wherein the expression vector is a plasmid.
41. The cDNA of claim 16 , wherein the polypeptide comprises:
an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:13 binds to human alpha-synuclein.
42. The cDNA of claim 16 , wherein the polypeptide comprises:
an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:10 binds to human alpha-synuclein.
43. The cDNA of claim 16 , wherein the polypeptide comprises:
an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO:10.
44. The cDNA of claim 16 , wherein the polypeptide comprises:
an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO:13.
45. The cDNA of claim 43 , which is in an expression vector.
46. The cDNA of claim 44 , which is in an expression vector.
47. The host cell of claim 17 , wherein:
the VH consists of the amino acid sequence set forth in SEQ ID NO:10; and
the VL consists of the amino acid sequence set forth in SEQ ID NO:13.
48. The host cell of claim 47 , wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.
49. A method for preparing an anti-human alpha-synuclein antibody, the method comprising:
culturing the host cell of claim 48 in a cell culture; and
isolating the anti-human alpha-synuclein antibody from the cell culture.
50. The method of claim 49 , further comprising formulating the anti-human alpha-synuclein antibody into a sterile pharmaceutical composition suitable for administration to a human subject.
51. The method of claim 50 , wherein the pharmaceutical composition is suitable for intravenous or subcutaneous administration.
52. The host cell of claim 17 , wherein the first expression vector is a plasmid, phage, or virus, and the second expression vector is a plasmid, phage, or virus.
53. The host cell of claim 17 , wherein the first expression vector is a retrovirus, and the second expression vector is a retrovirus.
54. The host cell of claim 17 , wherein the first expression vector comprises a first heterologous promoter operably linked to the first polynucleotide, and the second expression vector comprises a second heterologous promoter operably linked to the second polynucleotide.
55. The host cell of claim 54 , wherein the first heterologous promoter is a cytomegalovirus, simian virus 40, or retroviral promoter, and the second heterologous promoter is a cytomegalovirus, simian virus 40, or retroviral promoter.
56. The host cell of claim 54 , wherein the first heterologous promoter is the cytomegalovirus immediate early promoter and the second heterologous promoter is the cytomegalovirus immediate early promoter.
57. The host cell of claim 21 , wherein the expression vector is a plasmid, phage, or virus.
58. The host cell of claim 21 , wherein the expression vector is a retrovirus.
59. The host cell of claim 21 , wherein:
the VH consists of the amino acid sequence set forth in SEQ ID NO:10; and
the VL consists of the amino acid sequence set forth in SEQ ID NO:13.
60. The host cell of claim 59 , wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.
61. A method for preparing an anti-human alpha-synuclein antibody, the method comprising:
culturing the host cell of claim 60 in a cell culture; and
isolating the anti-human alpha-synuclein antibody from the cell culture.
62. The method of claim 61 , further comprising formulating the anti-human alpha-synuclein antibody into a sterile pharmaceutical composition suitable for administration to a human subject.
63. The method of claim 62 , wherein the pharmaceutical composition is suitable for intravenous or subcutaneous administration.
64. A polynucleotide linked to a heterologous nucleic acid, wherein the polynucleotide is selected from the group consisting of:
a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively, wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO:13 binds to human alpha-synuclein;
a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:10 binds to human alpha-synuclein;
a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:3, residues 50-66 of SEQ ID NO:3, and residues 99-110 of SEQ ID NO:3, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:7 binds to human alpha-synuclein;
a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:6, residues 49-55 of SEQ ID NO:6, and residues 88-98 of SEQ ID NO:6, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:4 binds to human alpha-synuclein;
a polynucleotide encoding an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:15, residues 50-66 of SEQ ID NO:15, and residues 99-112 of SEQ ID NO:15, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:19 binds to human alpha-synuclein;
a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 24-34 of SEQ ID NO:18, residues 50-56 of SEQ ID NO:18, and residues 89-96 of SEQ ID NO:18, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:16 binds to human alpha-synuclein; and
a polynucleotide encoding an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 24-34 of SEQ ID NO:21, residues 50-56 of SEQ ID NO:21, and residues 89-97 of SEQ ID NO:21, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:16 binds to human alpha-synuclein.
65. The polynucleotide of claim 64 , wherein:
the polynucleotide encodes an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 31-35 of SEQ ID NO:9, residues 50-68 of SEQ ID NO:9, and residues 101-102 of SEQ ID NO:9, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO:13 binds to human alpha-synuclein.
66. The polynucleotide of claim 64 , wherein:
the polynucleotide encodes an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in residues 23-33 of SEQ ID NO:12, residues 49-55 of SEQ ID NO:12, and residues 88-98 of SEQ ID NO:12, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO:10 binds to human alpha-synuclein.
67. The polynucleotide of claim 64 , wherein:
the polynucleotide encodes an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO:10.
68. The polynucleotide of claim 64 , wherein:
the polynucleotide encodes an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO:13.
69. The polynucleotide of claim 64 , wherein the heterologous nucleic acid is a regulatory element.
70. The polynucleotide of claim 69 , wherein the regulatory element is a promoter, an enhancer, a ribosome binding site, or a transcription terminator.
71. The polynucleotide of claim 64 , wherein the heterologous nucleic acid is a secretory signal peptide.
72. The polynucleotide of claim 71 , wherein the secretory signal peptide is a mammalian signal peptide.
73. The polynucleotide of claim 72 , wherein the mammalian signal peptide is from tissue plasminogen activator.
74. A host cell comprising the polynucleotide of claim 64 .
75. The host cell of claim 74 , wherein the host cell is a Chinese Hamster Ovary (CHO) cell.