Purification of type G botulinum neurotoxin and pharmaceutical compositions thereof
AssigneeWisconsin Alumni Research Foundation
InventorsEric A. Johnson, Hiroshi Sugiyama, Carl J. Malizio
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Abstract
The isolation and purification of type G botulinum neurotoxin and complexes thereof is disclosed. Compositions containing the type G neurotoxin and the preparation of a type G toxoid are also disclosed.
Claims4
A type G botulinum neurotoxin having a molecular weight of about 144,000 daltons as determined by gel filtration and SDS-PAGE, said neurotoxin having a specific toxicity of between 1.0.times.10.sup.7 and 5.0.times.10.sup.7 LD.sub.50 /mg protein wherein the neurotoxin was obtained from Clostridium botulinum grown under anaerobic conditions.
A method of obtaining natural type G botulinum neurotoxin which comprises
- (a) growing Clostridium botulinum strain 89 under anaerobic conditions in a suitable medium to obtain a culture broth having activity of between 3.0.times.10.sup.4 and 4.0.times.10.sup.4 LD.sub.50 /ml;
- and,
- (b) isolating the neurotoxin by adding yeast RNA and adjusting pH of the broth to about 3.4 to precipitate a toxin complex, adding RNAase and treating the precipitated toxin by DEAE Sephadex ion exchange purification and PAPTG affinity chromatography to obtain the isolated neurotoxin,
- wherein yield of the neurotoxin is between 9% and 11%.
A method of obtaining type G botulinum neurotoxin which comprises
- (a) growing Clostridium botulinum under anaerobic conditions in a suitable medium to obtain a culture broth having activity of >3.0.times.10.sup.4 and 4.0.times.10.sup.4 LD.sub.50 /ml;
- and,
- (b) isolating the neurotoxin by adding yeast RNA and adjusting pH of the broth to precipitate a toxin complex, adding RNAse and treating the precipitated toxin by ion exchange purification and affinity chromatography to obtain the isolated neurotoxin,
- wherein yield of the neurotoxin is between 9% and 11%.
Dependencies are inferred from the claim text.