Methods for purifying clostridial neurotoxin
AssigneeIpsen Biopharm Ltd.
InventorsGavin Stephen Hackett, Shilpa Palan, Dina Brady Anderson
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Abstract
A method for purifying a clostridial neurotoxin comprising contacting a cation exchange resin with a composition comprising a clostridial neurotoxin, wherein the contacting step is performed at at least pH 7.3, wherein the step of contacting a cation exchange resin with a composition comprising said clostridial neurotoxin occurs prior to conversion of the clostridial neurotoxin from a single chain form into a dichain form. Also provided are uses of a buffer having a pH value that is −1 pH unit or higher than the calculated pi of a clostridial neurotoxin, purification intermediates and clostridial neurotoxins obtainable by the invention, wherein the clostridial neurotoxin is in a single chain form.
Claims16
A method of producing the active di-chain form of a clostridial neurotoxin, the method comprising:
- (a) contacting a cation exchange resin with a composition comprising a single-chain clostridial neurotoxin, wherein the contacting step is performed at a pH of from 7.3 to 9.5;
- (b) separating the clostridial neurotoxin from the cation exchange resin and
- (c) contacting the separated clostridial neurotoxin with a protease that cleaves the polypeptide sequence of the clostridial neurotoxin under conditions suitable for the cleavage, thereby activating it to produce the active di-chain form of the clostridial neurotoxin,
- wherein the step (a) occurs before the step (b).
Dependencies are inferred from the claim text.