275706 Analysis of Metabolic Flux Rewiring in CHO Cells During Fed-Batch Culture

Tuesday, October 30, 2012: 8:30 AM
Washington (Westin )
Maciek R. Antoniewicz, Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE and Woo Suk Ahn, Chemical Engineering, University of Delaware, Newark, DE

Despite significant investments in research and development with the goal of improving understanding of CHO cells, relatively little is still known about intracellular metabolic fluxes in fed-batch cell cultures of CHO cells. In this work, the metabolism of CHO cells was studied in detail during a fed-batch fermentation. Metabolism was characterized by measured extracellular uptake and excretion rates of metabolites combined with a detailed metabolic network model for CHO cells. To obtain additional information 13C-tracers were applied and labeling of intracellular metabolites was measured using mass spectrometry (GC-MS). Fluxes were quantified using 13C-metabolic flux analysis. The flux results revealed significant rewiring of intracellular metabolic fluxes in the transition from growth phase to stationary phase, including changes in energy metabolism and redox metabolism. The results that we present provide a solid foundation for future studies of CHO cell metabolism for applications such as cell line development and medium optimization for high-titer production of recombinant proteins.

References

[1] Ahn WS, Antoniewicz MR. Metabolic flux analysis of CHO cells at growth and non-growth phases using isotopic tracers and mass spectrometry. Metab Eng, 13(5): 598-609, 2011

[2] Ahn WS, Antoniewicz MR. Towards dynamic metabolic flux analysis in CHO cell cultures. Biotechnol J, 7(1): 61-74, 2012


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