A Single Dynamic Metabolic Model Can Describe mAb Producing CHO Cell Batch and Fed-Batch Cultures on Different Culture Media

CHO cell culture high productivity relies on optimized culture medium management under fed-batch or perfused chemostat strategies enabling high cell densities. In this work, a dynamic metabolic model for CHO cells was further developed, calibrated and challenged using datasets obtained under four different culture conditions, including two batch and two fed-batch cultures comparing two different culture media. The recombinant CHO-DXB11 cell line producing the EG2-hFc monoclonal antibody was studied. Quantification of extracellular substrates and metabolites concentration, viable cell density, monoclonal antibody concentration and intracellular concentration of metabolite intermediates of glycolysis, pentose-phosphate and TCA cycle, as well as of energetic nucleotides, were obtained for model calibration. Results suggest that a single model structure with a single set of kinetic parameter values is efficient at simulating viable cell behavior in all cases under study, estimating the time course of measured and non-measured intracellular and extracellular metabolites. Model simulations also allowed performing dynamic metabolic flux analysis, showing that the culture media and the fed-batch strategies tested had little impact on flux distribution. This work thus paves the way to an in silico platform allowing to assess the performance of different culture media and fed-batch strategies.CHO细胞培养的高生产力依赖于补料分批（fed-batch）或灌注式恒化器（perfused chemostat）策略下的优化培养基管理，以实现高细胞密度。本研究进一步开发了针对CHO细胞的动态代谢模型，并通过四种不同培养条件下获取的数据集对模型进行校准与验证：其中包含2批批次培养（batch）与2批补料分批培养，且对比了两种不同的培养基。本研究以分泌EG2-hFc单克隆抗体的重组CHO-DXB11细胞株为研究对象，通过定量检测胞外底物与代谢物浓度、活细胞密度、单克隆抗体浓度，以及糖酵解、磷酸戊糖途径与三羧酸（TCA）循环的代谢中间产物胞内浓度，还有高能核苷酸浓度，为模型校准提供数据支撑。研究结果表明，采用单一模型结构与一组动力学参数值，即可有效模拟所有受试培养条件下的活细胞行为，准确估算实测与未实测的胞内、胞外代谢物的时间变化曲线。此外，通过模型模拟还可开展动态代谢流分析，结果显示受试培养基与补料分批策略对代谢流分布的影响极小。本研究为构建可评估不同培养基与补料分批策略性能的计算机模拟（in silico）平台奠定了基础。