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Background In CHO cell lines currently selected for the production of recombinant antibody, approximately 80% of the metabolized glucose is converted into lactic acid. These cells with a glycolytic phenotype exhibit significantly higher rates of proton production (extracellular acidification rate, ECAR) from lactate production than cells using oxidative phosphorylation (oxygen consumption rate, OCR). Therefore, shifts in the cell’s metabolism can be detected conveniently and dynamically through simultaneous detection of ECAR and OCR. Such measurements can characterize the metabolic programming of individual cell types and forecast the quality potential of their produced glycoproteins. In this study, we utilized an XF96 analyzer to measure glycolysis and mitochondrial respiration simultaneously, and in real-time. This allows one to determine the response of these two pathways to ATP demand, and indirectly, biosynthetic needs. A rapid screen was performed to determine the desired lactic acid production by exposing the cells to alternate sources of substrates, such as galactose or fructose. Specific metrics of the study included cell growth, product yield, and glycan profile. Higher titer, viable cell density, and viability along with glycol-similarity were observed for galactose and glutamine feeding strategies during the production phase. We believe this rapid, cell based metabolic screen that is label-free and non-invasive can be used to identify low lactic acid CHO mAb cell producers in both batch and fed-batch systems. This selection is accomplished without compromising clone productivity and product quality.

bmc proceedings

Analysis of glycolytic flux as a rapid screen to identify low lactate producing CHO cell lines with desirable monoclonal antibody yield and glycan profile