Interaction of cell culture process parameters for modulating mAb afucosylation. Issue 4 (4th February 2019)

• Record Type: Journal Article
• Authors: Nguyen Dang, Anh; Mun, Melissa; Rose, Christopher M.; Ahyow, Patrick; Meier, Angela; Sandoval, Wendy; Yuk, Inn H.
• Is Part Of: Biotechnology and bioengineering. Volume 116:Issue 4(2019)
• Page Start: 831
• Page End: 845
• Publication Date: 2019-02-04
• Subjects: afucosylation -- bioreactor scale‐up -- Chinese Hamster Ovary cell culture -- monoclonal antibody -- partial pressure of carbon dioxide -- proteomics; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.26908
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: The extent of afucosylation, which refers to the absence of core fucose on Fc glycans, can correlate positively with the antibody‐dependent cellular cytotoxicity (ADCC) activity of a monoclonal antibody (mAb). Therefore, it is important to maintain consistent afucosylation during cell culture process scale‐up in bioreactors for a mAb with ADCC activity. However, there is currently a lack of understanding about the impact of partial pressure of carbon dioxide (pCO2 )—a parameter that can vary with bioreactor scale—on afucosylation. Using a small‐scale (3 L) bioreactor model that can modulate pCO2 levels through modified configurations and gassing strategies, we identified three cell culture process parameters that influence afucosylation of a mAb produced by a recombinant Chinese Hamster Ovary (CHO) cell line: pCO2, media hold duration (at 37°C), and manganese. These three‐independent parameters demonstrated a synergistic effect on mAb afucosylation; increase in pCO2, media hold duration, and manganese consistently increased afucosylation. Our investigations into the underlying mechanisms through proteomic analysis indicated that the synergistic interactions downregulated pathways related to guanosine diphosphate‐fucose synthesis and fucosylation, and upregulated manganese transport into the CHO cells. These new findings highlight the importance of considering potential differences in culture environment and operations across bioreactor scales, and understanding the impact of their interactions on product quality. Abstract : The extent of afucosylation, which refers to the absence of core fucose on Fc glycans, can correlate positively with the antibody‐dependent cellular cytotoxicity (ADCC) activity of a monoclonal antibody (mAb). Therefore, it is important to maintain consistent afucosylation during cell culture process scale‐up in bioreactors for a mAb with ADCC activity. Using a small‐scale (3 L) bioreactor model that can modulate pCO2 levels through modified configurations and gassing strategies, we identified three cell culture process parameters that influence afucosylation of a mAb produced by a recombinant CHO cell line: pCO2, media hold, and manganese.