不同外源基因拷贝数背景CHO细胞的宿主细胞蛋白种类的研究  

Research of the host cell protein species with different exogenous gene backgrounds in CHO cells

作　　者：高侨 余垚[2] 蔡洁行 GAO Qiao;YU Yao;CAI Jie-xing(WuXi Biologics/State Key Laboratory of Genetic Engineering,School of Life Sciences,Fudan University;State Key Laboratory of Genetic Engineering,School of Life Sciences,Fudan University,Shanghai 200438,China;WuXi Biologics(CAI Jie-xing),Shanghai 200137,China)

机构地区：[1]上海药明生物技术有限公司/复旦大学生命科学院遗传工程国家重点实验室,200137 [2]复旦大学生命科学院遗传工程国家重点实验室,上海200438 [3]上海药明生物技术有限公司,200137

出　　处：《中国医药生物技术》2020年第2期171-177,共7页Chinese Medicinal Biotechnology

摘　　要：目的研究外源基因拷贝数、生产工艺、细胞多样性对宿主细胞蛋白表达的影响。方法构建空载体CHO K1细胞株,检测其外源基因拷贝数,根据拷贝数的低、中、高对细胞进行分类。使用不同的生产工艺测试低拷贝、中等拷贝和高拷贝的细胞群,观察其宿主细胞蛋白表达之间的差异。最后将三个拷贝数水平的细胞群进行混合,再生产,观察其混合表达的宿主细胞蛋白与单独生产时表达宿主细胞蛋白的差异。结果研究结果显示,不同拷贝数背景的细胞群之间表达宿主细胞蛋白相差6%~13%。相同的细胞进行不同的工艺生产时,也因为基础培养基和补料的不同而使宿主细胞蛋白的种类不同。细胞多样性增加并不能引起宿主细胞蛋白表达的种类增多。结论宿主细胞蛋白的表达与外源基因拷贝数、生产工艺以及细胞群的多样性有关。Objective This research was designed to explore the effect of exogenous gene copy number,process of production and cell diversity on host cell protein expression in CHO K1 cell pools.Methods We successfully developed three null cell lines of CHO K1 to detect the exogenous gene copy number levels,and classify the cells according to high,medium,low levels of gene copy number.Two processes were selected to evaluate three pools for the differential expression of HCPs.Finally,all three pools were mixed together and then the differential expression of HCPs between the mixed pools with unmixed ones were evaluated by process A.Results There were around 6%to 13%difference in HCPs expression among the pools with different exogenous gene copy number backgrounds.When we used the same cell line with different processes,the host cell proteins also showed differences from each other since the basic media and feed media were not the same.The addition of pool diversity didn’t lead to the increase of host cell protein species.Conclusion The host cell protein expression is influenced by exogenous gene copy number levels,process of the production and diversity of cells.

关 键 词：CHO细胞 宿主细胞蛋白 外源基因拷贝数 生产工艺 多样性 

分 类 号：Q78[生物学—分子生物学]