collagenase-as

CIzyme AS

$130.00

Description

CIzyme AS is an aseptically filled, lyophilized mixture of 60% purified class I (C1) and 40% purified class II (C2) collagenase from Clostridium histolyticum pre-mixed with neutral protease from Bacillus polymyxa.

Intended Use

CIzyme AS was developed to isolate adipose derived stromal cells (ADSC) from human lipoaspirate.1,2

Cat. # 005-1090
  • Manufactured for superior lot-to-lot enzyme consistency for improved performance consistency
  • Optimal enzyme activity (28-42 Wünsch Units) and neutral protease activity (220,000 – 340,000 NP units) per vial to dissociate approximately 100 grams of human adipose tissue.
  • Low contaminating endotoxin levels (<25 EU/OD)
  • 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.

Manufacturing Details

CIzyme AS collagenase is purified from culture supernatants of C. histolyticum that contain porcine gelatin and pancreatic enzymes derived from US and Canadian sources as the only animal derived material in the culture media. No animal derived products are used in the culture of B. polymyxa or in any step of the purification process for either enzyme.

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 AS 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 components used to prepare CIzyme AS. The Collagenase activity is characterized using both the traditional Wünsch (Pz-peptide) 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 substrate. The B. polymyxa neutral protease activity is relies on a FITC labeled BSA substrate. Focus on manufacture of products with consistent collagenase and neutral protease activity enables users to focus on other issues to improve their islet isolation protocols.

References

Product Documentation

Please 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.

Pack Insert

Safety Data Sheet

Specifications

Certificate of Animal Origin

General Product FAQ

Is this product intended for use in diagnostic or therapeutic procedures?

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.

What are application-specific formulations?

Application-specific formulations are pre-blended mixtures of purified collagenase and protease in optimal ratios for isolation of cells from a particular tissue and/or species.

Currently, our CIzyme™ Collagenase AS and CIzyme™ Collagenase RI are our only two application-specific formulations.

What is the best way to inactivate the enzymes when a digestion is complete?

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

How do I transition from a crude enzyme to a purified one?

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.

Why don’t you offer a formulation for my specific application?

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.

CIzyme AS FAQ

How should I store my enzyme?

This product is stable for at least two years from date of manufacture if stored unopened between -15 to – 25°C. Internal studies have shown the reconstituted enzyme is stable as a frozen solution between -15 to – 25°C for at least 3 months. Additional studies have shown the reconstituted enzyme was successfully frozen and thawed at least twice without apparent loss of potency as assessed by the CDA and NPA assays. The product is shipped on dry ice to provide the most stable conditions during shipment.