- Product consistency limits need for lot testing
- Consistent FALGPA activity units per bottle, with a fixed amount of protease activity and increasing amounts of collagenase activity
- Rapid dissolution, no clogged filters, no need for pre-centrifugation
- Use only what you need or reconstitute the entire bottle
- Low clostripain contamination
- Lot specific Certificate of Analysis provided with complete QC analysis results
DE Collagenase 20/200
VitaCyte’s defined enriched (DE) collagenase products set a new standard of consistency and enables assessment of a broader range of collagenase:protease ratios for cell isolation than found with current collagenase products. Each DE product contains a fixed amount of protease activity and increasing amounts of collagenase activity, simplifying collagenase selection and minimizing the need for lot qualification.
DE Collagenase 20 (100 mg pack size, Cat#011-1120) and DE Collagenase 200 (1 g pack size, Cat#011-1020) may be used in the following applications:
- Rat Hepatocytes
- Human Foreskin Fibroblasts
- Human Foreskin Keratinocytes
- Human Glioblastomas
- White lyophilized cake under vacuum in amber bottle sealed with butyl rubber stopper
- Stable for at least 2 years when stored at < + 4°C
- Lyophilized in a buffer containing calcium to ensure enzyme stability during initial reconstitution
DE Collagenase is an aseptically filled, lyophilized mixture of enriched Clostridium histolyticum collagenase and neutral protease from Bacillus polymyxa. Collagenase is purified from culture supernatants of C. histolyticum that contain porcine gelatin and pancreatic enzymes sterilized prior to bacterial fermentation. No animal derived products are used in the culture of B. polymyxa or in any step of the purification process for either enzyme. DE Collagenase contains a fixed amount of neutral protease activity (NP) units and variable amounts of enriched collagenase. DE Collagenase is prepared for research use only.
VitaCyte relies on several biochemical tools to characterize and ensure the consistency of components used to prepare DE Collagenase. The Collagenase activity is characterized using both the traditional Wünsch (Pz-peptide)1 substrate and a more recently developed fluorescent microplate collagen degrading activity (CDA) assays. The CDA relies on fluorecein isothiocyanate (FITC) labeled calf skin collagen fibrils as substrate2. The B. polymyxa neutral protease activity (NPA) is measured using a FITC labeled BSA substrate3.
Wünsch E and Heidrich H-G. (1963) Zur quantitativen bestimmung der kollagenase. Hoppe-Seyler’s Zeitschrift Physiologische Chemie 333, 149-151.
General Product FAQ
No. This product is for research use only. Guidance for use of reagents in clinical cell transplantation procedures is governed by local Institutional Review Boards and regional Health Authorities. This product is manufactured in accordance with the principles for clinical trial material outlined in ICH Q7a. The document control system in place is in alignment with FDA guidance for Phase I material. Document controls are in place to minimize the chances of cross-contamination.
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.