Therapeutic monoclonal antibodies that neutralize botulinum neurotoxins

1. An isolated antibody, or antigen-binding fragment thereof, that specifically binds to an epitope of Botulinum neurotoxin type A, wherein the antibody or antigen-binding fragment comprises:

a) a VL CDR1 comprising the amino acid sequence of SEQ ID NO:165;

b) a VL CDR2 comprising the amino acid sequence of SEQ ID NO:167;

c) a VL CDR3 comprising the amino acid sequence of SEQ ID NO:203; and

d) VH CDRs comprising:

i) a VH CDR1 comprising the amino acid sequence of SEQ ID NO:91;

ii) a VH CDR2 comprising the amino acid sequence of SEQ ID NO:93; and

ii) a VH CDR3 comprising the amino acid sequence of SEQ ID NO:130;

or

i) a VH CDR1 comprising the amino acid sequence of SEQ ID NO:95;

ii) a VH CDR2 comprising the amino acid sequence of SEQ ID NO:97; and

ii) a VH CDR3 comprising the amino acid sequence of SEQ ID NO:133.

2. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody is a single chain Fv (scFv).

3. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody is an IgG.

4. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody is a Fab.

5. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody is a (Fab′) 2 .

6. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody is a (scFv′) 2 .

7. The isolated antibody or antigen-binding fragment of claim 6 , wherein said antibody is a fusion protein of two scFv fragments.

8. A method of specifically binding a botulinum neurotoxin type A (BoNT/A), said method comprising contacting said botulinum neurotoxin type A with an anti-botulinum neurotoxin type A (anti-BoNT/A) antibody or antigen-fragment binding fragment according to claim 1 , said antibody or antigen-fragment binding fragment having a specificity and affinity such that it specifically binds to said botulinum neurotoxin type A.

9. The method of claim 8 , wherein said antibody binds both an A1 and A2 toxin.

10. The method of claim 8 , further comprising contacting said botulinum neurotoxin type A with a second anti-botulinum neurotoxin type A (anti-BoNT/A) antibody or antigen-fragment binding fragment according to claim 1 , said antibody or antigen-fragment binding fragment having a specificity and affinity such that it specifically binds to said botulinum neurotoxin type A, wherein said second anti-botulinum neurotoxin type A (anti-BoNT/A) antibody or antigen-fragment binding fragment binds to a different epitope than said first anti-botulinum neurotoxin type A (anti-BoNT/A) antibody or antigen-fragment binding fragment.

11. A composition comprising a plurality of anti-botulinum neurotoxin antibodies or antigen-binding fragments, wherein each antibody or antigen-binding fragment is specific for a different epitope of a botulinum neurotoxin, and wherein said plurality comprises an antibody of claim 1 .

12. The composition of claim 11 , wherein said composition comprises a first antibody or antigen-binding fragment that binds an A1 toxin and a second antibody or antigen-binding fragment that binds an A2 toxin.

13. A method of specifically binding a botulinum neurotoxin, said method comprising contacting said neurotoxin with the composition of claim 11 .

14. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody comprises a variable heavy (V H ) comprising complementarity determining regions comprising the amino acid sequence of V H CDR1 (SEQ ID NO: 95), V H CDR2 (SEQ ID NO: 97), and V H CDR3 (SEQ ID NO:133).

15. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody comprises a V H comprising complementarity determining regions comprising the amino acid sequence of V H CDR1 (SEQ ID NO: 91), V H CDR2 (SEQ ID NO: 93), and V H CDR3 (SEQ ID NO:130).

16. The isolated antibody or antigen-binding fragment of claim 1 , wherein said antibody or antigen-binding fragment comprises a full length light chain comprising framework regions comprising the amino acid sequence of V L framework 1 (SEQ ID NO: 164), of V L framework 2 (SEQ ID NO: 166), of V L framework 3 (SEQ ID NO: 202), and of V L framework 4 (SEQ ID NO: 204).

17. The isolated antibody or antigen-binding fragment of claim 16 , wherein said antibody or antigen-binding fragment comprises a variable heavy (V H ) comprising complementarity determining regions comprising the amino acid sequence of V H CDR1 (SEQ ID NO: 95), V H CDR2 (SEQ ID NO: 97), and V H CDR3 (SEQ ID NO:133).

18. The isolated antibody or antigen-binding fragment of claim 16 , wherein said antibody or antigen-binding fragment comprises a V H comprising complementarity determining regions comprising the amino acid sequence of V H CDR1 (SEQ ID NO: 91), V H CDR2 (SEQ ID NO: 93), and V H CDR3 (SEQ ID NO:130).

19. The isolated antibody or antigen-binding fragment of claim 17 , wherein said antibody or antigen-binding fragment comprises a V H comprising framework regions comprising the amino acid sequence of V H framework 1 (SEQ ID NO: 94), of V H framework 2 (SEQ ID NO: 96), of V H framework 3 (SEQ ID NO: 132), and of V H framework 4 (SEQ ID NO: 134).

20. The isolated antibody or antigen-binding fragment of claim 18 , wherein said antibody or antigen-binding fragment comprises a V H comprising framework regions comprising the amino acid sequence of V H framework 1 (SEQ ID NO: 90), of V H framework 2 (SEQ ID NO: 92), of V H framework 3 (SEQ ID NO: 129), and of V H framework 4 (SEQ ID NO: 131).

21. An isolated antibody, or antigen-binding fragment thereof, that specifically binds to an epitope of Botulinum neurotoxin type A, wherein the antibody comprises a full-length light chain comprising:

a V L CDR1 comprising the amino acid sequence of SEQ ID NO:165;

a V L CDR2 comprising the amino acid sequence of SEQ ID NO:167;

a V L CDR3 comprising the amino acid sequence of SEQ ID NO:203;

a V L framework 1 comprising the amino acid sequence of SEQ ID NO: 164;

a V L framework 2 comprising the amino acid sequence of SEQ ID NO:166;

a V L framework 3 comprising the amino acid sequence of SEQ ID NO: 202; and

a V L framework 4 comprising the amino acid sequence of SEQ ID NO: 204.

22. A method of detecting a Botulinum neurotoxin (BoNT) in a biological sample, the method comprising:

a) contacting the biological sample with the isolated antibody, or an antigen-binding fragment thereof, of claim 1 ; and

b) detecting binding, if any, of the antibody to an epitope present in the sample.