Antibodies that neutralize botulinum neurotoxins
This disclosure 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. 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 NYPMS (SEQ ID NO:520),
a V H CDR2 comprising the amino acid sequence of SLTASGDNTFYADSVKG (SEQ ID NO:522),
a V H CDR3 comprising the amino acid sequence of ALVGRYDISTGYYRPVMDS (SEQ ID NO:524); 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 RTSQGFTSALA (SEQ ID NO:770),
a V L CDR2 comprising the amino acid sequence of DASKLES (SEQ ID NO:772), and
a V L CDR3 comprising the amino acid sequence of QQSNSYPLT (SEQ ID NO:774).
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 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 thereof.
9. The method of claim 8 , wherein the method further comprises recovering the antibody or antigen-binding fragment thereof 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 thereof.
11. The method of claim 10 , wherein the method further comprises recovering the antibody or antigen-binding fragment thereof 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 the amino acid sequence of a variable heavy chain (VH) polypeptide comprising an amino acid of SEQ ID NO:23; and
b) a second isolated nucleic acid comprising a nucleotide sequence encoding the amino acid sequence of a variable light chain (VL) polypeptide comprising an amino acid sequence of SEQ ID NO:59.
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 thereof.
20. The method of claim 19 , wherein the method further comprises recovering the antibody or antigen-binding fragment thereof 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 thereof 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).