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US 7,452,697 B2Granted
Chromatographic method and system for purifying a botulinum toxin
AssigneeAllergan, Inc.
InventorsMingjiang Luo, Hui Xiang, Yao Yu, Stephen Donovan
Life of this patent
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Abstract
Chromatographic processes and systems for purifying a botulinum toxin from an APF fermentation medium.
Claims17
A substantially animal protein free (“APF”) process for purifying a biologically active botulinum toxin, the process comprising the steps of:
- (a) obtaining a fermentation medium which comprises a biologically active botulinum toxin, wherein the fermentation medium comprising a biologically active botulinum toxin results from a substantially, essentially or entirely APF fermentation process;
- (b) filtering the fermentation medium;
- (c) precipitating the botulinum toxin from the filtered fermentation medium to thereby obtain a precipitate;
- (d) extracting the botulinum toxin from the precipitate;
- (e) contacting a first chromatography column with the extracted botulinum toxin, wherein the first chromatography column is either a hydrophobic interaction chromatography column or an ion exchange chromatography column;
- (f) eluting the botulinum toxin from the first chromatography column;
- (g) contacting a second chromatography column with the botulinum toxin containing eluent from the first chromatography column, wherein the second chromatography column is either a hydrophobic interaction chromatography column or an ion exchange chromatography column;
- (h) eluting the botulinum toxin from the second chromatography column;
- (i) contacting a third chromatography column with the botulinum toxin containing eluent from the second chromatography column wherein the third chromatography column is either a mixed mode chromatography column or an ion exchange chromatography column, and;
- (j) eluting the botulinum toxin from the third chromatography column, thereby obtaining the purified biologically active botulinum toxin, wherein the first chromatography column, the second chromatography column, and the third chromatography column are different chromatography columns.
7 more elements
An APF process for purifying a biologically active botulinum toxin, the process comprising the steps of:
- (a) obtaining an APF fermentation medium which comprises a biologically active botulinum toxin, wherein the fermentation medium comprising a biologically active botulinum toxin results from an APF fermentation process;
- (b) filtering the APF fermentation medium;
- (c) precipitating the botulinum toxin from the filtered APF fermentation medium to thereby obtain a precipitate;
- (d) extracting the botulinum toxin from the precipitate;
- (e) contacting a first chromatography column with the extracted botulinum toxin, wherein the first chromatography column is either a hydrophobic interaction chromatography column or an ion exchange chromatography column;
- (f) eluting the botulinum toxin from the first chromatography column;
- (g) contacting a second chromatography column with the botulinum toxin containing eluent from the first chromatography column, wherein the second chromatography column is either a hydrophobic interaction chromatography column or an ion exchange chromatography column;
- (h) eluting the botulinum toxin from the second chromatography column;
- (i) contacting a third chromatography column with the botulinum toxin containing eluent from the second chromatography column wherein the third chromatography column is either a mixed mode chromatography column or an ion exchange chromatography column, and;
- (j) eluting the botulinum toxin from the third chromatography column, thereby obtaining the purified biologically active botulinum toxin, wherein the first chromatography column, the second chromatography column, and the third chromatography column are different chromatography columns.
7 more elements
An essentially animal protein free (“APP”) process for purifying a biologically active botulinum toxin, the process comprising the steps of:
- (a) obtaining a fermentation medium which comprises a biologically active botulinum toxin, wherein the fermentation medium comprising a biologically active botulinum toxin results from an essentially or entirely APF fermentation process;
- (b) filtering the fermentation medium;
- (c) precipitating the botulinum toxin from the filtered fermentation medium to thereby obtain a precipitate;
- (d) extracting the botulinum toxin from the precipitate;
- (e) contacting a first chromatography column with the extracted botulinum toxin, wherein the first chromatography column is either a hydrophobic interaction chromatography column or an ion exchange chromatography column;
- (f) eluting the botulinum toxin from the first chromatography column;
- (g) contacting a second chromatography column with the botulinum toxin containing eluent from the first chromatography column, wherein the second chromatography column is either a hydrophobic interaction chromatography column or an ion exchange chromatography column;
- (h) eluting the botulinum toxin from the second chromatography column;
- (i) contacting a third chromatography column with the botulinum toxin containing eluent from the second chromatography column wherein the third chromatography column is either a mixed mode chromatography column or an ion exchange chromatography column, and;
- (j) eluting the botulinum toxin from the third chromatography column, thereby obtaining the purified biologically active botulinum toxin, wherein the first chromatography column, the second chromatography column, and the third chromatography column are different chromatography columns.
7 more elements
Dependencies are inferred from the claim text.