Metabolic Profiling of CHO Cells during the Production of Biotherapeutics

Abstract

As indicated by an ever-increasing number of FDA approvals, biotherapeutics constitute powerful tools for the treatment of various diseases, with monoclonal antibodies (mAbs) accounting for more than 50% of newly approved drugs between 2014 and 2018 (Walsh, 2018). The pharmaceutical industry has made great progress in developing reliable and efficient bioproduction processes to meet the demand for recombinant mAbs. Mammalian cell lines are preferred for the production of functional, complex recombinant proteins including mAbs, with Chinese hamster ovary (CHO) cells being used in most instances. Despite significant advances in cell growth control for biologics manufacturing, cellular responses to environmental changes need to be understood in order to further improve productivity. Metabolomics offers a promising approach for developing suitable strategies to unlock the full potential of cellular production. This review summarizes key findings on catabolism and anabolism for each phase of cell growth (exponential growth, the stationary phase and decline) with a focus on the principal metabolic pathways (glycolysis, the pentose phosphate pathway and the tricarboxylic acid cycle) and the families of biomolecules that impact these circuities (nucleotides, amino acids, lipids and energy-rich metabolites).

Precision fermentation with mass spectrometry‐based spent media analysis

“…Malate, fumarate, citrate, and succinate are secreted in the exponential phase due to high TCA cycle activity for biomass production (Coulet et al, 2022). Kuang et al ( 2021) identified new inhibitory metabolites in CHO cell cultures that affect cell growth, protein titer, and mAb glycosylation, with the secretion of these metabolites being dependent on the cellular metabolic state (Supporting Information: Table S1) (Kuang et al, 2021).…”

A CHO stable pool production platform for rapid clinical development of trimeric SARS‐CoV‐2 spike subunit vaccine antigens

“…During subsequent decades, different groups, including nearly every major pharmaceutical company involved in biologics development, have been developing and optimizing their own CHO‐based expression platforms. The diversity of these platforms is notable, including differences in choice of CHO cell lineages, expression plasmid constructs, gene transfer and selection methods, culture media, and cell and process engineering strategies (Coulet et al, 2022; Donaldson et al, 2022; Durocher & Butler, 2009; Reyes et al, 2022; Wurm, 2013; Wurm & Wurm, 2017). However, a common feature has been the mandatory use of stable clonal cell lines for both clinical development and manufacturing, a result of several factors including longstanding regulatory guidelines (Stuible et al, 2018).…”

Addressing amino acid‐derived inhibitory metabolites and enhancing CHO cell culture performance through DOE‐guided media modifications

“…Robust cell growth, elevated viable cell densities, and high product titers can be achieved through proper utilization of glucose and amino acids and their efficient/effective conversion into energy, biomass, and bioproducts. Unfortunately, the disposition of nutrients to energy or useful metabolites does not always occur in cell culture processes (Coulet et al, 2022; Ritacco et al, 2018; Young, 2013). Alternatively, metabolic inefficiencies are manifested in part as the formation of pathway intermediates and their accumulation and secretion as by‐products in cell culture over time (Pereira et al, 2018).…”