In-depth Metabolic Characterization at Different Phases of Cell Culture with a Turnkey CE-ESI-HRMS Workflow

Understanding of cell culture metabolism in biologics manufacturing has increased tremendously in the past decade; however, the lack of feasible methods of high coverage, quantitative information on the interplay between nutrients and metabolites has hindered the efforts to fully characterize processes in the different biological phases. Several published models support cell growth phase but lack the kinetic information to be applicable to the phases that follow. We discuss a method for measuring hundreds of metabolites in dozens of pathways that streamlines sample preparation from spent media and cells, enables fast data-analysis times and supports quantitative analysis of key metabolites to gain far richer and more reliable data sources for process characterization, modelling and understanding. The method uses microchip CE/MS and incorporates stable isotope internal standards for both quantitation and migration time indexing. This work explores the information that can be obtained from the extracellular growth media and intracellular metabolites extracted from the CHO cells.

The poster was on display at Bioprocessing Summit, and ESACT #253 in 2024.