Industrial Chinese Hamster Ovary (CHO) cell strains used to produce biopharmaceuticals can grow in cultures with varying amounts of glutamine, and are characterized by significant changes in their metabolism over a typical fed-batch reactor. In order to understand CHO metabolism during exponential growth, and in particular the role of glutamine, we examined the utilization of 13C-glucose (U and 1,2), 13C-glutamine (U), and 13C-asparagine (U) for CHO cells cultures in limited and supplemented glutamine environments during the early and late exponential phase. Analysis of the intracellular free metabolites revealed differences in the allocation of amino acids to cellular physiology under different nutrient conditions. Likewise, enrichment of cell protein fixed in the biomass indicates different distribution of the 13C from the fed glucose, glutamine, and asparagine following changes in both the glutamine supplementation and time during the exponential phase. 13C metabolic flux analysis (13C MFA) of the data also indicated a change in the distribution of glucose, glutamine, and asparagine into glycolysis versus the TCA cycle for the early and late exponential phase. These findings will increase our understanding of CHO cellular metabolism in different nutrient environments and growth conditions in order to optimize both growth and ultimately protein production for CHO cells in fed-batch cultures.

Characterizing CHO cell metabolism in glutamine-limited and supplemented feeds during different stages of exponential growth

255th National Meeting (2018)

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