Combined in silico modeling and metabolomics analysis to characterize fed‐batch CHO cell culture

TLDR

The growing demand for recombinant therapeutic proteins drives the need to improve biopharmaceutical yield from mammalian cells, achievable only through a deep understanding of cellular function. The study aims to use combined metabolomics and in silico modeling to gain deeper insight into CHO fed‑batch culture mechanisms. The authors combined metabolomic profiling with in silico modeling of CHO cells to characterize metabolic behaviors across growth phases and identify growth‑limiting pathways such as oxidative stress and lipid depletion. Key metabolites in energy, glutathione, and glycerophospholipid pathways, distinct amino acid composition, and oxidative stress and lipid depletion were identified as growth‑limiting factors in CHO fed‑batch cultures. Published in Biotechnol.

Abstract

Abstract The increasing demand for recombinant therapeutic proteins highlights the need to constantly improve the efficiency and yield of these biopharmaceutical products from mammalian cells, which is fully achievable only through proper understanding of cellular functioning. Towards this end, the current study exploited a combined metabolomics and in silico modeling approach to gain a deeper insight into the cellular mechanisms of Chinese hamster ovary (CHO) fed‐batch cultures. Initially, extracellular and intracellular metabolite profiling analysis shortlisted key metabolites associated with cell growth limitation within the energy, glutathione, and glycerophospholipid pathways that have distinct changes at the exponential‐stationary transition phase of the cultures. In addition, biomass compositional analysis newly revealed different amino acid content in the CHO cells from other mammalian cells, indicating the significance of accurate protein composition data in metabolite balancing across required nutrient assimilation, metabolic utilization, and cell growth. Subsequent in silico modeling of CHO cells characterized internal metabolic behaviors attaining physiological changes during growth and non‐growth phases, thereby allowing us to explore relevant pathways to growth limitation and identify major growth‐limiting factors including the oxidative stress and depletion of lipid metabolites. Such key information on growth‐related mechanisms derived from the current approach can potentially guide the development of new strategies to enhance CHO culture performance. Biotechnol. Bioeng. 2012; 109:1415–1429. © 2012 Wiley Periodicals, Inc.