- Manufactured for superior lot-to-lot enzyme consistency for improved performance consistency
- Sufficient enzyme activity (>1,900 Wünsch Units) to release islets from average size adult human pancreata
- Purification process eliminates the majority contaminating protease activity including clostripain (trypsin-like activity)
- Low contaminating endotoxin levels (<10 EU/mg)
- Combine with choice of neutral protease for your established protocol
- Lot specific Certificate of Analysis provided with complete QC analysis results
Cat. # 001-1000
GMP Grade Collagenase HA is an aseptically prepared, lyophilized mixture of 60% purified Class I (C1) Collagenase and 40% purified Class II (C2) Collagenase from Clostridium histolyticum. This 2000 Wünsch Unit pack size can be used for isolating islets from human pancreata with the appropriate neutral protease.
- White lyophilized cake under vacuum in amber bottle sealed with butyl rubber stopper
- Highly purified collagenases for greater enzyme potency (>2.8 U/mg)
- Stable for at least 2 years when stored at -20°C
- Lyophilized in a buffer containing calcium to ensure enzymes stability during initial reconstitution
This product is purified from culture supernatants of C. histolyticum fermentation that contain porcine gelatin and pancreatic enzymes derived from US and Canadian sources. No bovine derived animal products are used in any step of the fermentation or purification process. 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 HA 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.
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 (FITC) labeled calfskin collagen fibrils as substrate.
The intact molecular form of purified C1 with two collagen binding domains (C1 116kDa) has approximately 10-fold higher CDA when compared the CDA found with same amount of purified C1 containing only one collagen binding domain (C1 100kDa) or intact C2 (C2 114kDa). A recent report in the literature describes the importance of intact C1 in successful human islet isolations.
Breite AG, Dwulet FE, McCarthy RC. 10th Annual Rachmiel Levine Diabetes and Obesity Symposium 2010 – Las Vegas, NV P#8 “Purification and Characterization of Clostridium histolyticum Neutral Protease used for Human Islet Isolation”
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.