Method for purification and activation of botulinum neurotoxin

A method of purifying Botulinum neurotoxin (BoNT) comprising the steps:

• a) contacting the BoNT to a compatible non-toxic non-hemagglutinin (NTNHA), under conditions appropriate for binding of the NTNHA to the BoNT to thereby form a NTNHA-BoNT complex.

A method of purifying Botulinum neurotoxin (BoNT) comprising:

• a) contacting a cleared cell extract comprising the BoNT to a glutathione coated matrix which has attached thereto a compatible non-toxic non-hemagglutinin (NTNHA) fused to glutathione-S-transferase, in a binding buffer with a pH of about 6 to thereby form a NTNHA-BoNT complex;
• b) washing the matrix with a wash buffer with a pH of about 6 to thereby remove unbound materials;
• c) eluting the BoNT from the matrix by contacting the matrix with an elution buffer that has a pH of ≥7.5 to thereby dissociate the BoNT from the NTNHA-BoNT complex.

A method of purifying a polypeptide comprising a receptor binding domain (Hc polypeptide) of Botulinum neurotoxin, comprising the steps:

• a) contacting a solution comprising the Hc polypeptide to a matrix which has attached thereto compatible non-toxic non-hemagglutinin (NTNHA), under conditions appropriate for binding of the NTNHA to the Hc-polypeptide to thereby form a NTNHA-Hc polypeptide complex;
• b) washing the matrix to thereby remove unbound materials;
• c) eluting the Hc polypeptide from the matrix by contacting the matrix with an aqueous solution that dissociates the Hc polypeptide from the NTNHA-Hc polypeptide complex.