Abstract
Chinese hamster ovary (CHO) cell expression systems are widely utilized for biologics manufacturing due to their efficient recombinant protein expression and human-like post-translational modifications. With an increasing demand for bio-therapeutic proteins, the development of high-yielding CHO cell lines and refining bioprocess parameters are key to achieving reliable, efficient, and cost-effective therapeutic protein production. Many lipidomic studies on CHO cells have highlighted the crucial roles of lipids in supporting cell growth and specific productivity, offering valuable insights for optimizing culture media and metabolic pathways. Among these lipid classes, sterols such as cholesterol and its downstream intermediates have been investigated in CHO cells, though their functions remain less thoroughly explored compared to overall lipid metabolism. In this study, comprehensive lipidomic profiling, including cholesterol and esterified cholesterol, was performed on two distinct basal media and feed conditions, which identified lipid composition changes that offer mechanistic insights into how culture conditions potentially influence membrane dynamics. These findings provide a foundation for future development of culture media and bioprocess control strategies that can ultimately improve yield and process consistency.