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Interaction of cell culture process parameters for modulating mAb afucosylation
Author(s) -
Nguyen Dang Anh,
Mun Melissa,
Rose Christopher M.,
Ahyow Patrick,
Meier Angela,
Sandoval Wendy,
Yuk Inn H.
Publication year - 2019
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26908
Subject(s) - chinese hamster ovary cell , bioreactor , fucosylation , fucose , antibody dependent cell mediated cytotoxicity , monoclonal antibody , cell culture , chemistry , glycosylation , microbiology and biotechnology , biochemistry , cytotoxicity , biology , antibody , in vitro , receptor , immunology , genetics , organic chemistry , glycoprotein
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 (pCO 2 )—a parameter that can vary with bioreactor scale—on afucosylation. Using a small‐scale (3 L) bioreactor model that can modulate pCO 2 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: pCO 2 , media hold duration (at 37°C), and manganese. These three‐independent parameters demonstrated a synergistic effect on mAb afucosylation; increase in pCO 2 , 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.