US 10,119,131 B2 · Compositions and methods for producing clostridial collagenases

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%.

Claims19

Dependent claims

A method for fermenting a clostripain-producing Clostridium histolyticum to obtain a mixture of collagenase I and collagenase II, comprising fermenting said clostripain-producing Clostridium histolyticum in medium comprising peptone, wherein the peptone is Oxoid VG100 Vegetable Peptone made from pea, or Oxoid VG200 Vegetable Peptone phosphate broth,

wherein said mixture of collagenase I and collagenase II is separately purified in the absence of added protease inhibitor and recombined to produce a drug product at least 95% pure as determined by reverse phase high performance liquid chromatography and contains no detectable clostripain activity using an enzymatic assay with N-a-Benzoyl-L-Arginine Ethyl Ester (BAEE) as substrate.

Dependencies are inferred from the claim text.

Compositions and methods for producing clostridial collagenases

Life of this patent

Compositions and methods for producing clostridial collagenases

Life of this patent

Abstract

Claims19