高密度接种和高密度冻存强化双特异性抗体细胞培养工艺  

Intensified bispecific antibody cell culture process using high seeding density and high cell density banking process

作　　者：张轩 吕明 丁晓然 谈彬 薛成 缪仕伟 ZHANG Xuan;LYU Ming;DING Xiao-ran;TAN Bin;XUE Cheng;MIAO Shi-wei(Hangzhou Sumgen Biotech Co.,Ltd,Hangzhou 310052,China)

机构地区：[1]杭州尚健生物技术有限公司,浙江杭州310052

出　　处：《生物技术》2023年第4期466-475,共10页Biotechnology

摘　　要：[目的]为了提高双特异性抗体生产效率,降低生产成本,对CHO细胞培养进行工艺强化研究。[方法]基于CHO细胞悬浮培养传统的补料批次生产工艺,应用高密度细胞冻存和N代高密度接种进行工艺强化,研究不同工艺对细胞生长、蛋白表达及关键质量属性的影响。[结果]将常规冻存密度1×10^(7)个/mL提高至1×10^(8)个/mL,在获得相当产量和可比产品质量的前提下,将细胞培养时间从23 d缩短至19 d;将初始接种活细胞密度从0.3×10^(6)~0.5×10^(6)cells/mL提高至3.0×10^(6)~6.0×10^(6)cells/mL;相同的培养周期内,在不影响蛋白质量的情况下最终蛋白产量从1.52 g/L提高至2.57 g/L。[结论]通过在2L生物反应器规模上对细胞培养工艺进行强化研究,缩短了4 d的细胞培养生产周期,降低了细胞污染的风险,在不影响蛋白质量的情况下,蛋白表达量提高了69%,降低了生产成本。[Objective]In order to increase antibody volumetric productivity,reduce the cost-of-goods(COG),it is necessary to intensify the Chinese hamster ovary(CHO)cell culture process.[Method]Based on conventional fed-batch culture of mammalian cells,high cell density banking process and N-generation high-density inoculation process were applied to investigate the influence of different process on cell growth,protein expression and critical quality attributes of antibodies.[Result]Increased conventional cell cryopreservation density from 1×10^(7) cells/mL to 1×10^(8) cells/mL could reduce the time required for culture from 23 d to 19 d with a comparable titer and product quality.The initial inoculated viable cell density was increased from conventional 0.3×10^(6)-0.5×10^(6) cells/mL to 3.0×10^(6)-6.0×10^(6) cells/mL,the final protein production was increased from 1.52 g/L to 2.57 g/L within the same culture time with the same product quality.[Conclusion]Through the intensified process strategy of Fed-batch process,it can shorten the production time of 4d,reduce the risk of cell contamination,improve protein expression was increased by 69%,reduce production costs.

关 键 词：双特异性抗体 CHO细胞 工艺强化 非灌注N-1种子培养 高密度接种 高密度冻存 补料批次培养 

分 类 号：Q815[生物学—生物工程]