Therapeutic monoclonal antibodies that neutralize botulinum neurotoxins
This invention provides antibodies that specifically bind to and typically neutralize botulinum neurotoxins (e.g., BoNT/A, BoNT/B, BoNT/E, etc.) and the epitopes bound by those antibodies. The antibodies and derivatives thereof and/or other antibodies that specifically bind to the neutralizing epitopes provided herein can be used to neutralize botulinum neurotoxin and are therefore also useful in the treatment of botulism.
1. An isolated antibody, or antigen-binding fragment thereof, that specifically binds an epitope of a Botulinum neurotoxin (BoNT), wherein the antibody or antigen-binding fragment comprises:
a V H CDR1 comprising the amino acid sequence of SEQ ID NO:125,
a V H CDR2 comprising the amino acid sequence of SEQ ID NO:127,
a V H CDR3 comprising the amino acid sequence of SEQ ID NO:129,
a V L CDR1 comprising the amino acid sequence of SEQ ID NO:229,
a V L CDR2 comprising the amino acid sequence of SEQ ID NO:231, and
a V L CDR3 comprising the amino acid sequence of SEQ ID NO:233; or
a V H CDR1 comprising the amino acid sequence of SEQ ID NO: 118,
a V H CDR2 comprising the amino acid sequence of SEQ ID NO: 120,
a V H CDR3 comprising the amino acid sequence of SEQ ID NO: 122,
a V L CDR1 comprising the amino acid sequence of SEQ ID NO: 222,
a V L CDR2 comprising the amino acid sequence of SEQ ID NO:224, and
a V L CDR3 comprising the amino acid sequence of SEQ ID NO:226.
2. A composition comprising the isolated antibody, or antigen-binding fragment thereof, of claim 1 and a pharmaceutically acceptable carrier.
3. The isolated antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody is a humanized antibody.
4. The isolated antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody is a human antibody.
5. The isolated antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody, or antigen-binding fragment thereof, is a single chain Fv (scFv), Fab, (Fab′) 2 or (scFv′) 2 .
6. The isolated antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody is an IgG.
7. A method of specifically binding a Botulinum neurotoxin in a mammal, said method comprising administering to said mammal the isolated antibody, or antigen-binding fragment thereof, of claim 1 .
8. A kit for specifically binding a Botulinum neurotoxin, said kit comprising:
the isolated antibody, or antigen-binding fragment thereof, of claim 1 ; and
a buffer.
9. The isolated antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody, or antigen-binding fragment thereof, comprises:
a V H CDR1 comprising the amino acid sequence of SEQ ID NO:118,
a V H CDR2 comprising the amino acid sequence of SEQ ID NO:120,
a V H CDR3 comprising the amino acid sequence of SEQ ID NO:122,
a V L CDR1 comprising the amino acid sequence of SEQ ID NO:222,
a V L CDR2 comprising the amino acid sequence of SEQ ID NO:224, and
a V L CDR3 comprising the amino acid sequence of SEQ ID NO:226.
10. The isolated antibody, or antigen-binding fragment thereof, of claim 9 , wherein said antibody comprises:
a heavy chain comprising the amino acid sequence of SEQ ID NO:45;
a light chain comprising the amino acid sequence of SEQ ID NO:60; or
a heavy chain comprising the amino acid sequence of SEQ ID NO:45 and a light chain comprising the amino acid sequence of SEQ ID NO:60.
11. A composition comprising the isolated antibody, or antigen-binding fragment thereof, of claim 10 and a pharmaceutically acceptable carrier.
12. The isolated antibody, or antigen-binding fragment thereof, of claim 10 , wherein said antibody, or antigen-binding fragment thereof, is a humanized antibody.
13. The isolated antibody, or antigen-binding fragment thereof, of claim 10 , wherein said antibody is a human antibody.
14. The isolated antibody, or antigen-binding fragment thereof, of claim 10 , wherein said antibody, or antigen-binding fragment thereof, is a single chain Fv (scFv), Fab, (Fab′) 2 or (scFv′) 2 .
15. The isolated antibody, or antigen-binding fragment thereof, of claim 10 , wherein said antibody is an IgG.
16. A method of specifically binding a Botulinum neurotoxin in a mammal, said method comprising administering to said mammal the isolated antibody, or antigen-binding fragment thereof, of claim 10 .
17. A kit for specifically binding a Botulinum neurotoxin, said kit comprising:
the isolated antibody, or antigen-binding fragment thereof, of claim 10 ; and
a buffer.
18. The isolated antibody, or antigen-binding fragment thereof, of claim 1 , wherein said antibody comprises:
a V H CDR1 comprising the amino acid sequence of SEQ ID NO:125,
a V H CDR2 comprising the amino acid sequence of SEQ ID NO:127,
a V H CDR3 comprising the amino acid sequence of SEQ ID NO:129,
a V L CDR1 comprising the amino acid sequence of SEQ ID NO:229,
a V L CDR2 comprising the amino acid sequence of SEQ ID NO:231, and
a V L CDR3 comprising the amino acid sequence of SEQ ID NO:233.
19. The isolated antibody, or antigen-binding fragment thereof, of claim 18 , wherein said antibody comprises:
a heavy chain comprising the amino acid sequence of SEQ ID NO:46;
a light chain comprising the amino acid sequence of SEQ ID NO:61; or
a heavy chain comprising the amino acid sequence of SEQ ID NO:46 and a light chain comprising the amino acid sequence of SEQ ID NO:61.
20. A composition comprising the isolated antibody, or antigen-binding fragment thereof, of claim 19 and a pharmaceutically acceptable carrier.
21. The isolated antibody, or antigen-binding fragment thereof, of claim 19 , wherein said antibody is a humanized antibody.
22. The isolated antibody, or antigen-binding fragment thereof, of claim 19 , wherein said antibody, or antigen-binding fragment thereof, is a human antibody.
23. The isolated antibody or antigen-binding fragment thereof, of claim 19 , wherein said antibody, or antigen-binding fragment thereof, is a single chain Fv (scFv), Fab, (Fab′) 2 or (scFv′) 2 .
24. The isolated antibody, or antigen-binding fragment thereof, of claim 19 , wherein said antibody is an IgG.
25. A method of specifically binding a Botulinum neurotoxin in a mammal, said method comprising administering to said mammal the isolated antibody, or antigen-binding fragment thereof, of claim 19 .
26. A kit for specifically binding a Botulinum neurotoxin, said kit comprising:
the isolated antibody, or antigen-binding fragment thereof, of claim 19 ; and
a buffer.
27. The isolated antibody, or antigen binding fragment of claim 10 , wherein said antibody comprises:
a heavy chain comprising the amino acid sequence of SEQ ID NO:45.
28. The isolated antibody, or antigen binding fragment of claim 10 , wherein said antibody comprises:
a light chain comprising the amino acid sequence of SEQ ID NO:60.
29. The isolated antibody, or antigen binding fragment of claim 10 , wherein said antibody comprises:
a heavy chain comprising the amino acid sequence of SEQ ID NO:45 and a light chain comprising the amino acid sequence of SEQ ID NO:60.
30. The isolated antibody, or antigen binding fragment of claim 19 , wherein said antibody comprises:
a heavy chain comprising the amino acid sequence of SEQ ID NO:46.
31. The isolated antibody, or antigen binding fragment of claim 19 , wherein said antibody comprises:
a light chain comprising the amino acid sequence of SEQ ID NO:61.
32. The isolated antibody, or antigen binding fragment of claim 19 , wherein said antibody comprises:
a heavy chain comprising the amino acid sequence of SEQ ID NO:46 and a light chain comprising the amino acid sequence of SEQ ID NO:61.
33. The isolated antibody, or antigen-binding fragment thereof, of claim 10 , wherein said antibody, or antigen-binding fragment thereof is a single chain Fv (scFv).