Animal product free system and process for purifying a botulinum toxin

Abstract

Chromatographic processes and systems for purifying a botulinum toxin from an APF fermentation medium.

I claim: 
     
         1 . A method for making a pharmaceutical composition essentially free of animal products in which the active ingredient is a botulinum toxin, the method comprising the steps of:
 (a) obtaining a biologically active botulinum toxin by
 (i) providing a fermentation medium free of animal products; 
 (ii) culturing a Clostridium botulinum in the fermentation medium under conditions which permit production of a botulinum toxin, and; 
 (iii) recovering a biologically active botulinum toxin from the fermentation medium; 
   (b) sequentially contacting a plurality of chromatography columns with an aqueous medium containing the recovered biologically active botulinum toxin; wherein the plurality of chromatography resins columns comprise at least a first chromatography column and a second chromatography column; wherein the first chromatography column is an ion exchange chromatography column, and the second chromatography column is a hydrophobic interaction chromatography column; wherein the biologically active botulinum toxin comprising a heavy chain and a light chain.   
     
     
         2 . The method of  claim 1 , wherein the plurality of chromatography resins columns further comprise a second ion exchange chromatography column. 
     
     
         3 . The method of  claim 1 , wherein the plurality of chromatography resins columns further comprise a mixed mode chromatography column. 
     
     
         4 . The method of  claim 1 , further comprising a filtration step comprising contacting the aqueous medium containing the biologically active botulinum toxin with a filtration filter. 
     
     
         5 . The method of  claim 1 , wherein the biologically active botulinum toxin is selected from the group consisting of botulinum toxin type A, B, C, D, E, F and G. 
     
     
         6 . The method of  claim 1 , wherein the biologically active botulinum toxin is a botulinum toxin type A.