Collagenases - Tissue Dissociation/Cell Isolation by Worthington Biochemical Corp.

One component of the complex is a hydrolytic enzyme which degrades the helical regions in native collagen preferentially at the Y-Gly bond in the sequence Pro-Y-Gly-Pro- where Y is most frequently a neutral amino acid. This cleavage yields products susceptible to further peptidase digestion.

Particular enzymatic profiles of each collagenase have been correlated with the tissues from which the cells for study were obtained (or with the uses to which the cells are put) and as a result of the correlations several types of crude collagenases have been established by Worthington:

• Type 1 crude collagenase has the original balance of collagenase, caseinase, clostripain and tryptic activities.
• Type 2 contains higher relative levels of protease activity particularly clostripain.
• Type 3 contains lowest levels of secondary proteases.
• Type 4 is designed to be especially low in tryptic activity to limit damage to membrane proteins and receptors.
• Purified collagenase, Code: CLSPA, contains minimal secondary proteolytic activities along with high collagenase activity.
• Animal Origin Free Types AFA, AFB and AFC collagenases are derived from cultures grown in medium completely devoid of animal based components and designed for bioprocessing applications where introduction of potential animal derived pathogens must be prevented. Levels of secondary proteases are similar to Types 1 and 2 collagenase.
• CLSAFA is the original AFA grade designed to have collagenase and secondary proteases similar to Types 1 and 2 collagenase.
• CLSAFB contains higher collagenase and caseinase activities than CLSAFA.
• CLSAFC has especially low tryptic activity similar to Type 4 collagenase.