Fast protein liquid chromatography of botulinum neurotoxin types A, B and E

Three antigenically different botulinum neurotoxins (NTs, relative molecular mass approximately 150,000), classically distinguished only by specific antisera, were for the first time chromatographically resolved. Mixed NTs eluted from a Mono-Q column in order of types E, A and B, and from Mono-S as B, E and A. Type A and B NTs were successfully chromatographed on the cation-exchange Mono-S column above their isoelectric points. Purification of type A and B NTs by automated liquid chromatography was also accomplished for the first time. Type A, B and E NTs were purified by application on an anion-exchange Mono-Q column, followed by use of a cation-exchange Mono-S column.