Despite intense efforts to improve Chinese hamster ovary (CHO) productivity, inherent cell culture stresses are unpredictable outcomes and can affect critical product quality attributes (CQAs). Common stressors in bioreactors are ammonia, lactate, and osmolarity, which are all metabolic byproducts that have been shown to adversely affect cell growth, protein productivity, and CQAs. In this study, industrially relevant levels these stressors were individually introduced to CHO cell cultures in an ambr® 250 system. Amino acid fluxes were quantified and glycosylation profiles for the monoclonal antibody, the main CQA, were characterized. As expected, the 30 mM ammonia-stress resulted in significantly decreased growth and poorer protein quality. In contrast, the 10 mM ammonia stress did not significantly affect growth until much later in the cultures. It was observed that the ammonia-stresses (10 mM and 30 mM ammonia) caused a dose-dependent response with respect to genome instability events. Conversely, the alanine flux profiles showed significant differences between the control and 10 mM ammonia-stressed cultures. A transcriptome approach was then used to discern the underlying metabolic shift in the cells due to the 10 mM ammonia-stress. Interestingly, initial ranges of osmolarity stress did not significantly affect cell growth. However, it has been shown that lactate buildup can affect cell productivity and growth. Combined, these results could lead to more accurate process and product quality predictions.

Utilization of the ambr®250 and a near real-time amino acid analyzer to understand the effects of common, industrially relevant mammalian cell culture stressors on process performance and product quality

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.