Increasing monoclonal antibody productivity from recombinant Chinese hamster ovary (CHO) cells continues to be a challenge in the biopharmaceutical industry. We developed an approach that utilizes flow cytometry methods for the detection of active capase-3 and cell cycle analysis as a guide to determine when to introduce anti-apoptotic agents that arrest cells in a more productive phase of the cell cycle. By frequent monitoring we were able to detect a sub-population of early membrane intact apoptotic cells with active caspase-3 prior to the reduction in membrane intact cell viability. Following detailed dose response studies, we introduced an effective concentration of Zn2+ ions at the appropriate time that increased membrane intact cell viability and induced arrest in G1 phase of the cell cycle. As a result, the Zn2+ supplemented cultures demonstrated a two-fold increase in cell specific productivity. We also characterized other bioreactor process strategies such as temperature shift by caspase-3 activation and cell cycle analysis to further increase protein productivity. Altogether we have gained greater insight into our bioreactor operations at the cellular level via this multiparametric analysis.

Strategy to enhance monoclonal antibody productivity in CHO cells with anti-apoptotic inhibitors based on flow cytometric analysis

253rd National Meeting (2017)

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