Animal product free system and process for purifying a botulinum toxin

An animal product free process for purifying a biologically active botulinum toxin, the process comprising the steps of:

• (a) preparing a botulinum toxin fermentation culture for passage over chromatography columns, wherein the fermentation culture is animal product free;
• (b) contacting a first chromatography column resin with prepared botulinum toxin fermentation culture, so as to permit capture of a botulinum toxin by the first column, wherein the first chromatography column resin utilizes a first separation mechanism selected from the group consisting of ion exchange, hydrophobic interaction, gel filtration and mixed mode mechanisms;
• (c) eluting the botulinum toxin from the first column;
• (d) loading a second column with eluent from the first column, wherein the second column interacts with eluent from the first column utilizing a second separation mechanism different from the first separation mechanism from the first column wherein the second separation mechanism is selected from the group consisting of ion exchange, hydrophobic interaction, gel filtration and mixed mode mechanisms;
• and
• (e) eluting the botulinum toxin from the second column, thereby obtaining a purified biologically active toxin.