表达TNFR-Fc融合蛋白的GS-CHO细胞动态流加培养过程的设计

Development of a fed-batch process for TNFR-Fc producing GS-CHO cells

摘要:

人肿瘤坏死因子受体Ⅱ-Fc融合蛋白在治疗风湿性、类风湿性关节炎方面拥有广阔的市场前景和巨大的经济价值。本实验以表达TNFR-Fc融合蛋白的GS-CHO细胞为研究对象，结合细胞生长代谢特性和动力学参数分析，以葡萄糖为关键控制参数，通过测定培养上清的葡萄糖浓度对培养过程中的葡萄糖消耗进行及时的预测，调整流加速率，形成了以满足细胞生长代谢需要为基本原则的动态流加培养过程设计模型。在此控制模型指导下，建立了高效的流加培养过程。使最大活细胞密度和最大融合蛋白浓度分别达9.4×106 cells/mL和207 mg/L，较批次培养分别提高了3.4倍和3倍。本研究所采用的研究方法和控制策略为优化GS-CHO细胞培养过程和TNFR-Fc融合蛋白成功迈向产业化奠定了基础。

Abstract:

TNFR-Fc is an important fusion protein that has great potential in therapeutic and diagnostic applications. We developed an efficient fed-batch process for GS-CHO cells to produce TNFR-Fc. The rationale of this fed-batch process relies on the supply of sufficient nutrients to meet the requirements of cell metabolism. The optimal feed medium was designed through ration design. A metabolically responsive feeding strategy was designed and dynamically adjusted based on the residual glucose concentration determined off-line. In this process, the maximal viable cell density and antibody concentration reached above 9.4×106 cells/mL and 207 mg/L, respectively. Compared with the batch process, the newly developed fed-batch process increased the cell yield by 3.4 fold and the final antibody concentration by 3 fold. This fed-batch process would therefore facilitate the production of therapeutic antibody by GS-CHO cells.