Traditional Crude or Enriched Collagenase
Minimally processed collagenase from Clostridium histolyticum culture supernatants; product first sold in the 1960s and contains collagenase and protease activities.
Purified Collagenase
Purified collagenase from other components in the C. histolyticum culture supernatants; products first sold in 1995 and require supplementation of the collagenases with protease enzymes.
Purified-Defined Collagenase
VitaCyte products that are similar to the purified collagenase but higher lot to lot consistency is ensured because of rigorous enzyme characterization.
Below compares different characteristics of traditional collagenase products to GMP Grade purified or purified-defined collagenase products manufactured by Nordmark, Roche, or VitaCyte.
Traditional Crude or Enriched Collagenase
Sigma or Worthington
Manufacturing Process:
Salt Precipitation
%
3-20% Purity
Lot Consistency:
Low
Lot Pre-Qualification:
Yes
Endotoxin Analysis:
GMP Grade:
Characterization
Peptidase:
FALGPA or Wunsch
CDA:
Mandl
Clostripain Activity:
Yes
General Protease Substrate
(Kinetic or Endpoint):
Casein
(Endpoint)
Anion Exchange
Chromotography:
No
Purified Collagenase
Nordmark or Roche
Manufacturing Process:
Column Chromatography
%
90% Purity
Lot Consistency:
Moderate to High
Lot Pre-Qualification:
Maybe
Endotoxin Analysis:
GMP Grade:
Characterization
Peptidase:
Wunsch
CDA:
Not Used
Clostripain Activity:
Yes
General Protease Substrate
(Kinetic or Endpoint):
DMC Casein
(Endpoint)
Anion Exchange
Chromotography:
Some Manufacturers
Purified-Defined Collagenase
VitaCyte
Manufacturing Process:
Column Chromatography
%
95% Purity
Lot Consistency:
High
Lot Pre-Qualification:
Not Needed
Endotoxin Analysis:
GMP Grade:
Characterization
Peptidase:
Wunsch
CDA:
Fluorescent-Kinetic
Clostripain Activity:
Yes
General Protease Substrate
(Kinetic or Endpoint):
FITC Human Serum Albumin
(Fluorescent-Kinetic)
Anion Exchange
Chromotography:
Yes
One important difference between the Purified vs Purified-Defined collagenase is the use of a fluorescent, kinetic microplate collagen degradation activity (CDA) assay. CDA is the only true measure of collagenase enzyme activity: degradation of native collagen. This assay has been rigorously characterized and each functional form of collagenase has a unique specific CDA. The function is defined by the ability of the collagenase to degrade native collagen.
The Mandl CDA is a research assay developed in the 1950s and has never been rigorously characterized for its precision or specificity. Moreover, the Wunsch assay preferentially detects the class II collagenase peptidase activity. This measurement is not a reliable measure of collagenase function since only the catalytic domain of the class II enzyme is required for Wunsch activity.