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.

1 - 6 . (canceled) 
     
     
         7 . A process for obtaining purified  botulinum  toxin type A comprising:
 (a) Preparing a  botulinum  toxin fermentation culture for passage over chromatography columns, wherein the fermentation culture is animal product free;   (b) contacting a hydrophobic interaction chromatography (HIC) column resin with the prepared  botulinum  toxin fermentation culture;   (c) washing the HIC column resin to remove impurities from said  botulinum  toxin A;   (d) eluting the  botulinum  toxin from the HIC column resin;   (e) contacting an anion exchange column resin with the eluant from the HIC column resin;   (f) eluting the  botulinum  toxin from the anion exchange column resin;   (g) contacting a cation exchange column resin with the eluant from the anion exchange column; and   (h) eluting the  botulinum  toxin from the cation exchange column, wherein the eluant comprises purified  botulinum  toxin type A.   
     
     
         8 . The process of  claim 7 , wherein said purified  botulinum  toxin type A comprises a purified, approximately 150 kDa  botulinum  toxin type A molecule. 
     
     
         9 . The process of  claim 7 , wherein said purified  botulinum  toxin type A comprises a purified, approximately 900 kDa  botulinum  toxin type A toxin complex. 
     
     
         10 . A purified  botulinum  toxin type A that is essentially free of animal product, made by the process plurality of chromatography steps. 
     
     
         11 . The purified  botulinum  toxin type A of  claim 10 , wherein said  botulinum  toxin type A product comprises a purified, approximately 150 kDa  botulinum  toxin type A molecule. 
     
     
         12 . The purified  botulinum  toxin type A of  claim 10 , wherein said  botulinum  toxin type A product comprises a purified, approximately 900 kDa  botulinum  toxin type A toxin complex. 
     
     
         13 . A fermentation media for culturing  Clostridium botulinum  that is essentially free of animal product, said fermentation media comprising about 3-6 wt % hydrolyzed soy protein. 
     
     
         14 . The fermentation media of  claim 13 , further comprising about 0-3% wt % yeast extract. 
     
     
         15 . The fermentation media of  claim 13 , further comprising about 1 wt % glucose. 
     
     
         16 . The fermentation media of  claim 13 , wherein said hydrolyzed soy protein is about 6 wt %. 
     
     
         17 . The fermentation media of  claim 13 , wherein said yeast extract is about 1 wt %. 
     
     
         18 . The fermentation media of  claim 13 , wherein the pH of said fermentation medium is about 5.0-5.5. 
     
     
         19 . A process for making a formulated  botulinum  toxin type A product that is essentially free of animal product comprising:
 (a) obtaining purified  botulinum  toxin type A from a fermentation culture that is essentially free of animal products; and   (b) combining said purified  botulinum  toxin type A from said sample with at least one pharmaceutically acceptable excipient that is essentially free of animal products to provide a formulated  botulinum  toxin type A product that is essentially free of animal products.   
     
     
         20 . The method of  claim 19 , wherein said purified  botulinum  toxin type A comprises a purified, approximately 150 kDa  botulinum  toxin type A molecule. 
     
     
         21 . The method of  claim 19 , wherein said purified  botulinum  toxin type A comprises a purified, approximately 900 kDa  botulinum  toxin type A toxin complex.