Loss of productivity during the industrial production of monoclonal antibodies is sometimes observed in recombinant Chinese Hamster Ovary cells. This work highlights a case study demonstrating upstream process development efforts to address challenges associated with cell line instability and low production yield. Two molecular mechanisms known to cause production instability are loss of recombinant gene copies after subculturing over many cell generations and epigenetic gene silencing via DNA methylation. Genetic characterization of our cell line revealed no loss of gene copies and that addition of a DNA methylation inhibitor, 5-Aza-2'-deoxycytidine (Decitabine), had a negligible effect suggesting that the change in phenotypic performance is not attributed to this typical genetic mechanism. Instead both clone stability and productivity are heavily dependent on passaging and production media, along with the feeds used as part of a fed-batch process. Evaluation of multiple commercial and in-house media and feeds led to the identification of an optimized combination, which resulted in minimal loss of stability over 60 population doublings while increasing titer in the fed-batch production cultures by 4-fold. Further development revealed that additional metal supplementation during the production step corrected the undesired metabolic profiles and phenotypes associated with cell age. Overall, we were able to develop a robust and manufacturable seed expansion and production scheme with stable and improved cell culture performance while maintaining comparable product quality attributes.

Modulating cell line stability through media and process selection for a monoclonal antibody

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.