Glucose withdrawal-induced Chinese hamster ovary cells apoptosis and its potential role in process robustness assessment

Abstract Establishing a robust process has always been one of the main goals of cell culture process development. To achieve this goal, glucose needs to be maintained at a low concentration to reduce lactate production, the main metabolic by-product. Keeping glucose low for long periods of time is a big challenge which can lead to glucose withdrawal. This study found that prolonged glucose withdrawal led to CHO cells apoptosis and the tolerance time of CHO cells to glucose withdrawal differed with the process conditions and the culture phases. A robust cell culture process allowed CHO cells to tolerate glucose withdrawal longer time, suggesting that the robustness of the process may be judged by the time duration for which CHO cells tolerated glucose withdrawal. Currently, assessment of process robustness is often time-consuming, labor-intensive, and material-intensive by process characterization studies. Therefore a simple and time-saving method is highly needed for the biopharmaceutical industry. The aim of this study was to investigate glucose withdrawal-induced apoptosis and develop a simple and time saving strategy for assessing the robustness of cell culture processes. The strategy we present may be used to expedite the assessment of process robustness to obtain a robust cell culture process for other biologics.