Patent ID: 8129139

Claim:
A substantially animal product free (APF) chromatographic process for obtaining a biologically active botulinum neurotoxin type A complex, the process comprising the following sequential steps: (a) culturing Clostridium botulinum bacteria in a substantially APF culture medium; (b) fermenting Clostridium botulinum bacteria from the culture medium in about 2 L to about 75 L of a substantially APF fermentation medium, wherein at least one of the culture medium and the fermentation medium includes a vegetable protein; (c) harvesting the fermentation medium by removing cellular debris present in the fermentation medium; (d) concentrating the harvested fermentation medium by filtration; (e) diluting the concentrated fermentation medium by adding a buffer; (f) a first contacting step in which the diluted harvested fermentation medium is contacted with an anion exchange media so that the biologically active botulinum neurotoxin becomes captured with the anion exchange media; (g) eluting the captured botulinum neurotoxin from the anion exchange media to thereby obtain a first eluent; (h) a second contacting step in which the first eluent is contacted with a cation exchange media to remove impurities from the first eluent, to thereby obtain a second eluent; (i) processing the second eluent by diafiltration; and (j) filtering the processed second eluent, thereby obtaining biologically active botulinum neurotoxin type A complex using a substantially APF chromatographic process, wherein the botulinum neurotoxin type A complex obtained has a potency of 2.0 times 10 7 to about 6.0 times 10 7 units/mg of botulinum neurotoxin type A complex.