Abstract
Neurotoxins of Clostridium botulinum are needed in basic neurologic research, but as therapeutic agent for certain neuro-muscular disorders like strabism as well. A method for the production and purification of botulinum neurotoxins C and D is reported using a two-step hollow-fiber cross flow filtration and a newly developed chromatographic purification procedure. Hollow-fiber filtration proved to be a rapid and safe concentration and pre-purification step, which can easily be scaled up. The chromatographic purification included hydrophobic interaction, anion exchange and size exclusion chromatography runs. Botulinum neurotoxins C and D could be recovered with an overall yield of 12.6% and 10.6%, respectively. A specific toxicity of 1.86×10 7 minimal lethal dose mg −1 (type C) and 5.26×10 7 minimal lethal dose mg −1 (type D) was determined in the mouse bioassay.
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