Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, People's Republic of China.
Jecho Laboratories, Inc., 7320 Executive Way, Frederick, MD, 21704, USA.
Bioprocess Biosyst Eng. 2019 Aug;42(8):1263-1271. doi: 10.1007/s00449-019-02124-7. Epub 2019 Apr 13.
In recent years, an increasing number of defucosylated therapeutic antibodies have been applied in clinical practices due to their better efficacy compared to fucosylated counterparts. The establishment of stable and clonal manufacturing cell lines is the basis of therapeutic antibodies production. Bioprocess development of a new cell line is necessary for its future applications in the biopharmaceutical industry. We engineered a stable cell line expressing defucosylated anti-HER2 antibody based on an established α-1,6-fucosyltransferase (FUT8) gene knockout CHO-S cell line. The optimization of medium and feed was evaluated in a small-scale culture system. Then the optimal medium and feed were scaled up in a bioreactor system. After fed-batch culture over 13 days, we evaluated the cell growth, antibody yield, glycan compositions and bioactivities. The production of anti-HER2 antibody from the FUT8 gene knockout CHO-S cells in the bioreactor increased by 37% compared to the shake flask system. The N-glycan profile of the produced antibody was consistent between the bioreactor and shake flask system. The antibody-dependent cellular cytotoxicity activity of the defucosylated antibody increased 14-fold compared to the wild-type antibody, which was the same as our previous results. The results of our bioprocess development demonstrated that the engineered cell line could be developed to a biopharmaceutical industrial cell line.
近年来，由于去岩藻糖基化治疗性抗体的疗效优于岩藻糖化抗体，越来越多的去岩藻糖基化治疗性抗体被应用于临床实践。建立稳定的和克隆的生产细胞系是治疗性抗体生产的基础。新细胞系的生物工艺开发是其未来在生物制药行业应用的必要条件。我们基于已建立的α-1,6-岩藻糖基转移酶（FUT8）基因敲除 CHO-S 细胞系，构建了表达去岩藻糖基化抗 HER2 抗体的稳定细胞系。在小规模培养系统中评估了培养基和补料的优化。然后，在生物反应器系统中放大最佳培养基和补料。经过 13 天的分批补料培养，我们评估了细胞生长、抗体产量、聚糖组成和生物活性。与摇瓶系统相比，FUT8 基因敲除 CHO-S 细胞在生物反应器中的抗 HER2 抗体产量增加了 37%。生物反应器和摇瓶系统中产生的抗体的 N-糖基化谱一致。与野生型抗体相比，去岩藻糖基化抗体的抗体依赖细胞毒性活性增加了 14 倍，与我们之前的结果相同。我们的生物工艺开发结果表明，该工程细胞系可以开发成为生物制药工业细胞系。
