Use VitaCyte’s Collagenase-Protease Products in Place of Liberase™
The enzyme composition for the largest pack sizes of GMP Grade and non GMP Grade Roche Liberase products are found in the table below. The Clostridium histolyticum collagenase used in these products are a 60:40 mixture of purified, intact class I (C1):class II (C2) collagenase as was used in the original Liberase product line.(1)
Thermolysin and Dispase are included in the Liberase products MNP-S, DL, DH, TL, TM, TH. For the MTF and T-Flex kits, Thermolysin is included as separate 15 mg bottles.
The mass of Thermolysin used in Liberase formulas were changed in 2008. This difference is based on the extinction coefficients (A280/mg protein) used to calculate the Thermolysin protein concentration. For the Modified Liberase, the value was 1.76, whereas for the Original Liberase the value was 1.1.
To calculate equivalent enzyme activities between Roche and VitaCyte’s purified Thermolysin products:
- Each mg of the Original Liberase or VitaCyte Thermolysin product is equivalent to 0.625 mg of the Modified Liberase Thermolysin product.
- Each mg of Modified Liberase Thermolysin is equivalent to 1.6 mg of the Original Liberase or VitaCyte Thermolysin product.
The shaded far right column indicates the amount of VitaCyte Thermolysin or Dispase™ enzyme you would use to achieve a similar neutral protease activity as found in the Roche Liberase products.
Modified Liberase Product Line2008 to Present |
Catalog #(R) = Roche (S) = Sigma |
Target mg Purified Collagenase per BottleLarge Pack Size |
Neutral Protease |
Ave Target Proteasemg/bottle |
Target mg VitaCyte’s Purified Proteases |
| Liberase MTF*GMP Grade | 05339880001 (R) | 537 | Thermolysin: Three separate 15 mg bottles | ≈ 24 in each bottle | |
| Liberase MNP-SGMP Grade | 06297790001 (R) | 4.7 | Thermolysin | 2.2 | ≈ 3.5 |
| Liberase MNP-S | 05578566001 (R) | 35 | Thermolysin | 16.9 | ≈ 27 |
| Liberase T-Flex | 05989132001 (R) | 537 | Thermolysin: Two separate 15 mg bottle | ≈ 24 in each bottle | |
| Liberase DL | 5466202001 (S) | 50 | Dispase | ? | ? |
| Liberase DH | 5401089001 (S) | 50 | Dispase | ? | ? |
| Liberase TL | 5401020001 (S) | 5.0 | Thermolysin | 0.23 | ≈ 0.37 |
| Liberase TM | 5401127001 (S) | 50 | Thermolysin | 23.3 | ≈ 37.3 |
| Liberase TH | 5401151001 (S) | 50 | Thermolysin | 46.6 | ≈ 74.6 |
VitaCyte’s products are listed below. Collagenase HA is equivalent to the Roche purified collagenase used in all the products above. The only differences between VitaCyte’s purified Collagenase HA and the Roche purified collagenase are the subclone derived from the parent strain of Clostridium histolyticum used to manufacture collagenase for the original Liberase products and the methods used to purify the collagenases. The collagenase C1:C2 ratios found in the Collagenase HA and Liberase MTF products are the same (60:40 C1:C2).
Product |
Catalog Number |
Pack Size |
Approximate Massmg |
Manufacturing Grade |
| Collagenase HA | 001-1000 | 2000 Wunsch U | 450 | GMP Grade |
| Collagenase HA | 001-1050 | 200 Wunsch U | 45 | GMP Grade |
| Collagenase MA | 001-2020 | 1100 Wunsch U | 250 | GMP Grade |
| Collagenase MA | 001-2030 | 2.5 million CDA U | 100 | GMP Grade |
| Collagenase Gold | 011-1060 | 1 g | 440 | Research Use Only |
| Collagenase Gold Plus | 011-2000 | ≥ 1500 FALGPA U | 450 | GMP Grade |
| Thermolysin | 002-3000 | 6 mg | 6 | AOF, GMP Grade |
| BP Protease | 003-1000 | 1.1 million NPA U | 8 | AOF, GMP Grade |
| BP Protease | 003-2000 | 2.3 million NPA U | 16 | AOF, GMP Grade |
| rCollagenase HI | 001-4010 | 1600 Wunsch U | 380 | AOF, GMP Grade |
| CIzyme RI | 005-1030 | 375K CDA U/ 275K NP U | 16 | Research Use Only |
| CIzyme AS | 005-1090 | 35 Wunsch U 280K NP U | 9 | GMP Grade |
VitaCyte has a broad line of purified collagenase products. In addition to selling Collagenase HA (high activity), VitaCyte sells Collagenase MA (moderate activity), a purified 60:40 C1:C2 mixture where intact and truncated class I purified collagenases are mixed with intact class II collagenase. Collagenase MA has a lower specific collagen degradation activity than found in Collagenase HA (55,000 vs 25,000 CDA U/mg enzyme).
Collagenase Gold, is a minimally processed C. histolyticum collagenase that has a C1:C2 ratio of about 75:25.(2) About 85% of this product contains intact C1 and C2 with a purity > 95%. The remaining proteins are degraded byproducts of C. histolyticum collagenase. Collagenase Gold is as effective as higher priced, purified collagenase enzymes for human islet isolation.(2, 3) The Collagenase Gold Plus product is a GMP Grade version of the same product.
rCollagenase HI is a GMP Grade, animal origin free, recombinant C. histolyticum collagenase designed for human islet isolation. The enzyme formulation was derived from a design of experiment study performed at the University of Minnesota to determine the effect of C1 and C2 collagenase dose on human islet isolation.(4) The formulation was the mid-point of the four different doses used in the study. Recent work at the University of Alberta in Edmonton has shown that human islets isolated with rCollagenase HI and BP Protease gave significantly higher stimulation indices in glucose stimulated insulin release studies, higher recovery of islets after culture that translated to higher numbers of islets used in the transplant procedure when compared to islets isolated with natural purified collagenase (MTF kit or Collagenase HA-thermolysin).(5)
BP Protease is a GMP Grade, Dispase equivalent enzyme. It has a specific neutral protease activity about 70% higher than other purified Dispase products sold today. The higher specific activity reflects the intact Dispase enzyme whereas other products undergo auto-catalysis which reduced the enzymes specific activity.
The CIzyme RI and CIzyme AS are products specifically designed for rodent islet isolation and human stromal vascular cell isolation from adipose tissue collected by liposuction.
For any technical questions, Contact Us or in the US call 888-664-2687.
References
1. McCarthy RC, Green ML, Dwulet FE. Evolution of enzyme requirements for human islet isolation. OBM Transplantation. 2018; 2. Available from: http://www.lidsen.com/journals/transplantation/transplantation-02-04-024.
2. Loganathan G, Hughes MG, Szot GL, Smith KE, Hussain A, Collins DR, et al. Low cost, enriched collagenase-purified protease enzyme mixtrues successfully used for human islet isolation. OBM Transplantation. 2019; 3. Available from https://www.lidsen.com/journals/transplantation/transplantation-03-02-064.
3. Khiatah B, Tucker A, Chen KT, Perez R, Bilbao S, Valiente L, et al. Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata. Islets. 2018; 10(2):51-9.
4. Balamurugan AN, Green ML, Breite AG, Loganathan G, Wilhelm JJ, Tweed B, et al. Identifying Effective Enzyme Activity Targets for Recombinant Class I and Class II Collagenase for Successful Human Islet Isolation. Transplantation Direct. 2016; 2(1):e54.
5. O’Gorman D, Kin T, Rosichuk S, Richer Brad, Zhai, W, Moriarty J, et al. 306.4: Evaluation of a Low Dose Recombinant Collagenase and BP Protease for Clinical Islet Transplantation. 2021; 105:S19-S20.