Compositions and methods for producing clostridial collagenases
AssigneeBioSpecifics Technologies Corp.
InventorsThomas L. WEGMAN, Bo YU
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Abstract
The present invention provides a method for producing a drug product comprising a combination of highly purified collagenase I and collagenase II from Clostridium histolyticum. The method utilizes an improved medium for the cultivation of Clostridium histolyticum which includes a non-meat-derived (i.e., non-mammalian) peptone or vegetable peptone. The method includes one or more of: (1) reducing glucose content in the meat-free or vegetable-derived media; and (2) increasing the salt concentration in the meat-free or vegetable-derived media. Also provided is a drug product which includes collagenase I and collagenase II at an optimized fixed mass ratio, and which has a purity of greater than at least 95%.
Claims25
A method for preparing a drug product comprising isolated and purified collagenase I and collagenase II having the sequence of Clostridium histolyticum collagenase I and collagenase II, said method comprising
- a) fermenting Clostridium histolyticum in an animal-derived material-free medium to obtain a crude collagenase mixture, wherein said medium is treated by:
- (1) decreasing the glucose content in the animal derived material-free medium;
- or (2) increasing the salt concentration in the animal derived material-free medium;
- b) separately purifying collagenase I and collagenase II from the crude collagenase mixture;
- and
- c) recombining the purified collagenase I and collagenase II at an optimized fixed mass ratio;
- wherein said drug product is at least 95% pure and substantially free of any contaminating proteases.
A medium for the fermentation of Clostridium histolyticum comprising:
- a. phytone peptone;
- b. yeast extract;
- and
- c. less than about 5 g/L glucose;
- wherein the pH of said medium is 7.5-7.9.
A medium for the fermentation of Clostridium histolyticum comprising:
- d. phytone peptone;
- e. yeast extract;
- and
- f. greater than about 5 g/L salt;
- wherein the pH of said medium is 7.5-7.9.
Dependencies are inferred from the claim text.