Use Of Roche Recombinant Trypsin For Cell Culture Applications

By Dr. Andrea Normann, Dr. Manuela Poignee-Heger, and Dr. Peter Hloch

Since 2004, Roche has produced recombinant trypsin (Roche Recombinant Trypsin) using a yeast strain, Pichia pastoris. The manufacturing process follows Good Manufacturing Practice (GMP) guidelines and uses no animal-derived products. The enzyme's production is regularly audited by a major insulin manufacturer because it is often used in creating Active Pharmaceutical Ingredients (APIs).

As the demand for replacing native enzymes with recombinant ones in human-use products grows, this application note demonstrates how Roche Recombinant Trypsin can be used in cell culture. We specifically highlight its use for detaching adherent cells, particularly those used in vaccine production.

To achieve this, we first optimized the reaction buffer and enzyme concentration to detach sensitive MRC-5 cells without causing damage. We then optimized the protocol for other common cell lines used in vaccine production, such as FRhK-4, Vero, MDCK, and CHO cells. Finally, we determined the thermal stability of the diluted enzyme solution.