- Entirely animal origin free
- Manufactured for superior lot-to-lot enzyme consistency for improved performance consistency
- Sufficient enzyme activity to release islets from average size adult human pancreata
- Lot specific Certificate of Analysis provided with complete QC analysis results
AOF rCollagenase HI
(Cat. # 001-4010)
rHI is an aseptically prepared, lyophilized mixture of purified recombinant collagenase C1 (ColG) and purified recombinant collagenase C2 (ColH) from engineered E. coli.
rHI is manufactured to provide the optimal collagenase activity for recovering islets from human pancreata when combined with purified BP Protease.
- White lyophilized cake under vacuum in amber bottle sealed with butyl rubber stopper
- Highly purified recombinant collagenases blended to minimize the amount of activity required for maximum islet recovery
- 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 E. coli cells grown in entirely animal origin free media components. Standard chromatography techniques are used to purify recombinant collagenase from separate strains to near homogeneity. After assessing enzyme potency for each rC1 and rC2, the recombinant collagenases are sterile filtered, aseptically blended and aseptically dispensed into depyrogenated vials followed by lyophilization. When fully dry, the rHI is sealed under vacuum. Representative samples are reconstituted and analyzed by a variety of quality control tests and compared to established specifications before release into inventory.
General Product FAQ
Thermolysin and BP Protease are entirely animal origin free.
A porcine gelatin peptone is used during the fermentation step of natural collagenase production. While this animal sourced material is required to stimulate collagenase production from the biosynthesis from the organism Clostridium histolyticum during anaerobic fermentation, it represents a very low adventitious agent risk as the media is sterilized at 121°C for 30 minutes prior to inoculation, which has been shown to inactivate many viruses.
The gelatin is removed during the purification process and no animal sourced materials are used in the remainder of the process.
Additional manufacturing details are available in product insert literature for each product.
There are two components to inactivation of the digestive enzymes commonly used for tissue dissociation. To inactivate protease activity, a serum-containing media or buffer is most effective. Buffers or media containing human serum albumin (HSA) are ineffective in suppressing protease activity, even at very high concentrations of protein (>10%). However, serum-containing buffers do not inhibit collagenase activity. The most effective method for inactivating collagenase activity is reducing temperature. Collagenases will lose about 80% their maximal activity at 26°C. Thus, introduction of an ice-cold, serum- containing media should virtually eliminate all collagenolytic and proteolytic activity of an enzyme mixture.
General Collagenase FAQ
VitaCyte’s technical service team has a significant amount of experience in transitioning from crude enzyme products to the more defined purified ones and is glad to provide you with assistance. If you can provide us details of the cell of interest; the enzyme product that is currently being used for the isolation; the concentrations (or activities) being targeted; volumes utilized; and, a certificate of analysis for the particular lot you would like to mimic; we can provide some reasonable guidance to which product(s) and what target activities best fit your application. Our defined enriched (DE) collagenase products are specifically designed to encompass the broad range of activities found in the crude enzyme products and to make the transition to a highly purified product seamless.
In developing an Application Specific Formulation, we generally rely on individuals considered “expert in the art” to ß-test potential formulations and guide us in the refinement of the final composition. These formulations are usually engineered from existing protocols and crude enzyme targets. Once a formulation has been identified and deemed acceptable by at least a couple independent labs, it may be included in our product portfolio. We are always eager to assist you with virtually any request for a custom blend so do not hesitate to contact us to discuss your application.