Antibodies for botulinum neurotoxins
The present disclosure provides antibodies that specifically bind to botulinum neurotoxins (e.g., BoNT/A, BoNT/B, BoNT/C, BoNT/D, BoNT/E, BoNT/F, BoNT/G, etc.) and the epitopes bound by those antibodies. The antibodies and derivatives thereof 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. A composition comprising:
a) a first isolated nucleic acid comprising a nucleotide sequence encoding a variable heavy chain (V H ) polypeptide comprising:
a V H CDR1 comprising the amino acid sequence of residues 31-35 of NHWMQ (SEQ ID NO:68),
a V H CDR2 comprising the amino acid sequence of residues 50-66 of RINDSGRSTSYAGFVKG (SEQ ID NO:68),
a V H CDR3 comprising the amino acid sequence of residues 99-114 of ETWEVLGHLGYEVLDH (SEQ ID NO:68); and
b) a second isolated nucleic acid comprising a nucleotide sequence encoding a variable light chain (V L ) polypeptide comprising:
a V L CDR1 comprising the amino acid sequence of residues 24-34 of RASQGISSYLA (SEQ ID NO:205),
a V L CDR2 comprising the amino acid sequence of residues 50-56 of AASTLQS (SEQ ID NO:205), and
a V L CDR3 comprising the amino acid sequence of residues 89-97 of QQLNSYPRT (SEQ ID NO:205).
2. The composition of claim 1 , wherein the first and second isolated nucleic acids are on the same nucleic acid constructs.
3. An expression vector comprising the first and second isolated nucleic acids of claim 2 .
4. A cell comprising the expression vector of claim 3 .
5. The composition of claim 1 , wherein the first and second isolated nucleic acids are on different nucleic acid constructs.
6. A cell comprising the first and second isolated nucleic acids of claim 5 .
7. The cell of claim 6 , wherein the said different nucleic acid constructs are expression vectors.
8. A method of making an antibody, or antigen-binding fragment thereof, the method comprising culturing the cell of claim 6 under conditions suitable for the cell to express the antibody or antigen-binding fragment.
9. The method of claim 8 , wherein the method further comprises recovering the antibody or antigen-binding fragment from the cell culture.
10. A method of making an antibody, or antigen-binding fragment thereof, the method comprising culturing the cell of claim 4 under conditions suitable for the cell to express the antibody or antigen-binding fragment.
11. The method of claim 10 , wherein the method further comprises recovering the antibody or antigen-binding fragment from the cell culture.
12. The method of claim 11 , wherein the antibody, or antigen-binding fragment thereof, is a single chain variable fragment (ScFv).
13. A composition comprising:
a) a first isolated nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of a variable heavy chain (V H ) polypeptide comprising an amino acid of SEQ ID NO:68; and
b) a second isolated nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of a variable light chain (V L ) polypeptide comprising an amino acid sequence of SEQ ID NO:205.
14. The composition of claim 13 , wherein the first and second isolated nucleic acids are on the same nucleic acid constructs.
15. A expression vector comprising the first and second isolated nucleic acids of claim 14 .
16. The composition of claim 13 , wherein the first and second isolated nucleic acids are on different nucleic acid constructs.
17. A cell comprising the first and second isolated nucleic acids of claim 16 .
18. The cell of claim 17 , wherein said different nucleic acid constructs are expression vectors.
19. A method of making an antibody or antigen-binding fragment thereof, the method comprising culturing the cell of claim 17 under conditions suitable for the cell to express the antibody or antigen-binding fragment.
20. The method of claim 19 , wherein the method further comprises recovering the antibody or antigen-binding fragment from the cell culture.
21. A cell comprising the expression vector of claim 15 .
22. A method of making an antibody, or antigen-binding fragment thereof, the method comprising culturing the cell of claim 21 under conditions suitable for the cell to express the antibody, or antigen-binding fragment thereof.
23. The method of claim 22 , wherein the method further comprises recovering the antibody or antigen-binding fragment from the cell culture.
24. The method of claim 23 , wherein the antibody, or antigen-binding fragment thereof, is a single chain variable fragment (ScFv).