Collagenase MA
(Cat. # 001-2030)

$252.00

GMP Grade Collagenase MA is an aseptically filled, lyophilized protein mixture containing 60% purified Class I (C1) and 40% purified Class II (C2) collagenase from Clostidium histolyticum. This 2.5 Million CDA Unit pack size can be used for isolating human hepatocytes with the appropriate neutral protease.

Cat. # 001-2030 Categories: , ,

 

  • Manufactured for superior lot-to-lot enzyme consistency for improved performance consistency
  • Low contaminating endotoxin levels (<10 EU/mg)
  • Combine with your choice of neutral protease for your established protocol
  • Lot specific Certificate of Analysis provided with complete QC analysis results
  • Lyophilized in a buffer containing at least 0.1 mM Ca2+ calcium to ensure optimal enzyme activity and stability
  • Highly purified collagenases for greater enzyme potency (>2.8 U/mg)
  • Unopened product stable for at least 2 years when stored between -15 and -25 °C
  • Lyophilized in a buffer containing at least 0.1 mM Ca2+ calcium to ensure optimal enzyme activity and stability
  • Highly purified collagenases for greater enzyme potency (>2.8 U/mg)
  • Unopened product stable for at least 2 years when stored between -15 and -25 °C

Collagenase MA is purified from culture supernatants of C. histolyticum that contain porcine gelatin and pancreatic enzyme derived from US and Canadian sources. No bovine derived animal products are used in any step of the fermentation or purification process. Collagenase MA is formulated to contain a sufficient amount of collagen degrading activity (CDA) units for the islet isolation application. The purification process ensures that a negligible amount of trypsin-like activity (i.e., contaminating clostripain) is present in the product.

Standard chromatography techniques are used to purify the collagenase. After assessing enzyme potency, the collagenase is aseptically dispensed into depyrogenated vials and lyophilized. When dry, the Collagenase MA vials are sealed under vacuum. A representative sample from the batch is reconstituted and analyzed by a variety of quality control tests. Results are compared to established specifications before release into inventory.

 

Suggested Assays

VitaCyte relies on several biochemical tools to characterize and ensure the consistency of Collagenase MA. The Pz-peptide substrate (Wünsch Assay) has historically been used to characterize collagenase activity. While this assay has advantages in terms of reproducibility and historical precedence, it also has several limitations. The Wünsch Assay is strongly biased towards C2 and is not sensitive to the different molecular forms of C1.

In addition, this activity assesses the catalytic activity of the enzyme and not its ability to degrade native collagen. Degraded C2 without a collagen binding domain is as active as intact C2, potentially providing misleading information about the quality of the enzyme. The limitations of the Wünsch assay led us to develop a fluorescent microplate CDA using fluorecein isothiocyanate labeled calf skin collagen fibrils as substrate.

The intact molecular form of purified C1 with two collagen binding domains(C1116kDa) has approximately 10-fold higher CDA when compared the CDA found with same amount of purified C1 containing only one collagen binding domain (C1100kDa) or intact C2 (C2114kDa). Collagenase MA is manufactured with both C1116kDa and C1100kDa to provide the optimal collagenase activity for the isolation of islets from NHP. Focus on manufacture of products with consistent collagenase activity enables users to focus on other issues to improve their islet isolation protocols.

References

  1. Green ML, Nonweiler KK, Breite AG, et al. Midwest Islet Club 3rd Annual Conference 2010 – Indianapolis, IN “Optimizing the Parameters for Porcine Islet Isolation: A Two-Year History”

Find the Pack Insert, Safety Data Sheet, Specifications, and Certificate of Animal Origin in the Product Documentation page.

Note: To obtain Certificates of Analysis for our products, please contact VitaCyte. Have your product lot number readily available to help us expedite your request.