Interaction of cell culture process parameters for modulating mAb afucosylation

Area of Science:

• Biotechnology
• Bioprocess Engineering
• Glycobiology

Background:

• Afucosylation of Fc glycans on monoclonal antibodies (mAbs) correlates with enhanced antibody-dependent cellular cytotoxicity (ADCC).
• Consistent afucosylation is vital for mAb efficacy, especially during process scale-up in bioreactors.
• The influence of partial pressure of carbon dioxide (pCO2) on mAb afucosylation during scale-up remains poorly understood.

Purpose of the Study:

• To investigate the impact of pCO2 and other cell culture parameters on mAb afucosylation.
• To identify key process parameters influencing afucosylation in recombinant Chinese Hamster Ovary (CHO) cell cultures.
• To elucidate the underlying mechanisms of afucosylation modulation.

Main Methods:

• Utilized a small-scale (3L) bioreactor model with controlled pCO2 levels.
• Modulated pCO2, media hold duration, and manganese concentrations.
• Employed proteomic analysis to investigate molecular mechanisms.

Main Results:

• Identified pCO2, media hold duration, and manganese as key parameters influencing mAb afucosylation.
• Demonstrated a synergistic effect where increased levels of these three parameters consistently elevated afucosylation.
• Proteomic analysis revealed downregulation of fucose synthesis pathways and upregulation of manganese transport.

Conclusions:

• pCO2, media hold duration, and manganese exhibit synergistic effects on mAb afucosylation.
• Understanding these interactions is critical for controlling product quality during bioreactor scale-up.
• These findings provide insights into optimizing cell culture processes for enhanced mAb efficacy.