Cell Suppliers
The focus of end-users who work within this segment is to provide high viability, functional cell populations to research or biopharmaceutical scientists who use these cells for their experimental or developmental work. The fresh or cryopreserved cells used for these applications are isolated and processed by commercial firms, organ procurement organizations, or core laboratories within academic universities or research institutes.
End-uses within these organizations strongly desire consistent collagenase-protease enzyme mixtures so that few adjustments are required when a new lot of a collagenase-protease product is used for isolating cells. Continuity of product supply is also a critical issue because failure to manufacture cells because of enzyme supply compromises the organization’s mission or profitability. Cost control of the isolation procedures is always at the forefront of these organizations.
VitaCyte’s Support for Manufacturing Cells
VitaCyte addresses three concerns end-users may have when evaluating different enzyme suppliers to support products used for manufacturing cells.
Consistency
Continuity of Product Supply
Enzyme Cost
Consistency
All purified collagenase products are > 95% pure. The analyses or enzyme activity assays described below assess the GMP-grade collagenase products.
Analytical anion exchange chromatographic analysis for purity and identity including the detection of degraded class I collagenase
Collagenase peptidase activity using FALGPA or Pz-peptide (Wünsch) assays
Collagen degradation activity (CDA) using a fluorescent, kinetic microtiter plate assay and fluorescein isothiocyanate (FITC) conjugated to purified type 1 calf collagen fibrils
Unactivated (i.e., trypsin-like activity) and activated (clostripain) BAEE activity
Endotoxin expressed as EU/A280
The analytical anion exchange analysis and Wünsch assay use the same methods described in US Pharmacopeia monographs <89.1> and <89.2> for class I (C1) and class II (C2) C. histolyticum collagenases, respectively. The collagen degradation activity (CDA) has been extensively characterized. Collagen degradation activity is the only activity measurement of collagenase function
The between lot coefficients of variation of enzyme activity assays independent lots of manufactured product is typically ≤ 15%. The primary assays used for these analyses is a kinetic fluorescent microtiter plate assay for measuring collagen degradation activity or neutral protease activity and a Wünsch assay to measure class II collagenase activity
VitaCyte also offers several GMP Grade purified neutral proteases essential for enzyme-mediated cell isolation.
Characteristics of VitaCyte’s GMP Grade Neutral Proteases
| Thermolysin | BP Protease | AM Protease | Clostripain | |
|---|---|---|---|---|
| Source | Bacillus thermoproteolyticus | Paenibacillus polymyxa | Aspergillus melleus | C. histolyticum |
| Assay Substrate | FITC-HSA | FITC-HSA | FITC-HSA | BAEE Peptide |
| Specific Activity U/mg Protein | 260,000 NP U/mg | 140,000 NP U/mg | 154,000 NP U/mg | 80 BAEE U/mg |
| Between Lot CVs | 7.7%, n=5 | 11.5%, n=15 | 14.3%, n=8 | 12.5%, n=5 |
Continuity of Product Supply
VitaCyte ensures a stable supply chain by maintaining a 6- to 9-month raw material reserve. In addition, a robust supplier qualification process provides continuity of raw materials for manufacturing and testing processes.
Enzyme Cost
Enzyme costs are controlled by offering the broadest choice of purified-defined collagenase enzymes. The end-user can select the appropriate collagenase product based on their needs to reduce enzyme costs, adopt an established cell isolation protocol using VitaCyte enzymes, or use animal-origin-free products for manufacturing therapeutic cells to avoid the risks of adventitious agents and related discussions with regulatory officials.
Collagenase Gold
$
Animal Origin Free
Defined C1:C2 Ratios
Endotoxin Testing
GMP Grade
Collagenase Gold Plus
$$
Animal Origin Free
Defined C1:C2 Ratios
Endotoxin Testing
GMP Grade
AOF rCollagenase HI*
$$$$
Animal Origin Free
Defined C1:C2 Ratios
Endotoxin Testing
GMP Grade
*AOF animal-origin-free; all other products are generated from C. histolyticum culture media that contains porcine gelatin peptone
The differences in price between the different product categories reflect the manufacturing process and extent of testing required to meet customer expectations that balance product characteristics with product application. The collagenase Gold products contain the C1 and C2 collagenase ratios generated by C. histolyticum in the fermentation procedure, whereas C1 and C2 are separately purified and re-blended in the Collagenase HA, Collagenase MA, and rCollagenase HI products. Collagenase Gold is a non-GMP, low-cost purified collagenase because it is not as extensively characterized as the Collagenase Gold Plus (identical to Collagenase Gold but with no excipient), Collagenase HA, collagenase MA and rCollagenase HI.
If you need additional technical information for any VitaCyte product or any information about the use of collagenase protease mixtures in enzyme-mediated cell isolation, Contact Us.
