Consumption of nutrients and production of metabolites lead to environmental changes over the course of cell-culture processes. Cell metabolism both drives and is affected by these changes that are often accompanied with decreased productivity and product quality. The general metabolic characteristics of cells lines derived from the same host cell are generally similar, however, differences in the metabolic profiles in culture are commonly seen. We hypothesize that the difference in cell line metabolic performance is the result of variability in the composition of metabolic machinery and the topology of regulatory network, and that the effect of those variations can be captured and predicted by a mechanistic metabolic model adapted for a particular cell line. In this work, we systematically characterize the transcriptomic, metabolic and N-Glycosylation profiles of two CHO cell lines to integrate with a mechanistic kinetic metabolic model for the metabolic characterization of each cell line. The model incorporates mechanistic reactions of glycolysis, the pentose phosphate pathway, the citric acid cycle, redox balance, and anaplerosis and cataplerosis between compartments. The model also depicts the regulatory effects of signaling pathways such as AKT and AMPK. Transcriptional abundance of metabolic enzymes, including various isozymes, are used for specification of the model.<br/><br/>A detailed survey of nutrient and metabolite dynamics was performed. Stable isotope labeling experiments were performed to determine carbon flux at major branches of metabolism. The dynamics of flux distribution upon metabolic shifts from lactate production to lactate consumption were determined, and the model used to predict the dynamics of metabolic shift. Protein quality attributes, particularly N-glycosylation, present measurable changes over the course of the fed-batch process, which we evaluate in the context of metabolic behavior. The system engineering approach employed in this study is applicable to the rational design of bioprocesses through process and product characterization and metabolic modeling.

Integration of a mechanistic kinetic model in metabolic characterization of cell lines

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.