Patent ID: 7452697

Claim:
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.