- Manufactured for superior lot-to-lot enzyme consistency for improved performance consistency
- Low contaminating endotoxin levels (<50 EU/OD)
- Lot specific Certificate of Analysis provided with complete QC analysis results
AOF BP Protease
(Cat. # 003-2000)
BP Protease is an affordable, aseptically dispensed, lyophilized protein containing purified neutral protease from Bacillus polymyxa.1 Sold in 2.3 million NP Unit pack size. This is the same enzyme as BP Protease (Cat# 003-1000) but now available in a pack size for those that often use 2 bottles per isolation.
BP Protease is a Dispase™ equivalent, neutral protease enzyme, a superb product to isolate human epithelial skin cells from tissue or after in vitro culture1,2. This enzyme is also a component of the DE Collagenase products and is used in enzyme mixtures that also contain Collagenase HA and Collagenase MA.
These combinations have been successfully used to isolate islets2, hepatocytes, and adipose derived stromal cells (ADSC), and are versatile and flexible enough to use for many other cell isolation applications.
- White lyophilized cake under vacuum in amber bottle sealed with butyl rubber stopper
- Vial contains 2.3 million Neutral Protease (NP) units, with total NP activity >1.8 million NP Units
- Stable for at least 2 years when stored at < -20 °C
This product is purified from culture supernatants of of B. polymyxa using column chromatography. No animal derived components were used in the culture or the subsequent purification steps.
After assessing enzyme potency, the protease is aseptically dispensed into depyrogenated vials and lyophilized. When dry, the BP Protease 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 specific neutral protease activity of purified BP Protease is determined by the FITC-BSA substrate NP3. The specific activity is used to provide a total target number of NP units.
McCarthy RC, Dwulet FE, Breite AG, Green M. World Stem Cell Summit 2014 – San Antonio, TX “A Cost-effective, Improved Preparation of Purified Paenbacillus polymyxa Neutral Protease (Dispase® Equivalent Enzyme)”
Balamurugan AN, Green M, Breite A, Loganathan G, Wilhem J, Vargova L, Lockridge A, Hering B, Dwulet F, McCarthy R. 2014 World Transplant Congress – San Francisco, CA “Human Pancreas Digestion and Islet Yields When Using With Purified P. polymyxa Protease Enzyme Mixtures”
BP Protease FAQ
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 6 months without loss of potency. Additional internal studies have shown the enzyme can go through a freeze-thaw cycle at least once without loss of potency by neutral protease (NP) assay.
The “BP” in BP Protease stands for “Bacillus polymyxa,” from which this purified neutral protease was derived.
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 Protease FAQ
Yes. Selection of a protease in many cases is simply due to the historical precedence of the given cell isolation protocol being followed; however, that is not to say a different one would not work. The isolation of human islets is a perfect example in which thermolysin, BP Protease (a Dispase equivalent), and C. histolyticum neutral protease have all been successfully used. Generally, bacterial or fungal sources of protease are selected due to their broader specificity. Substitution of one protease for another may not be a seamless transition, however, as the kinetics of digestion can be dramatically different among the various general proteases. For guidance in selecting a protease, contact VitaCyte’s technical support team.