高效治疗性抗体CHO细胞株的构建方法及其评价  

Construction and evaluation of Chinese hamster ovary cell lines with a high therapeutic antibody

作　　者：刘治影 罗顺 吕芊锐 LIU Zhi-ying;LUO Shun;LYU Qian-rui(College of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012;Drug Research Institute of Yangtze Detla,Nantong Jiangsu 226133;Thousand Oaks Biopharma,Nantong Jiangsu 221600)

机构地区：[1]安徽中医药大学药学院,合肥230012 [2]南通市海门长三角药物高等研究院,江苏南通226133 [3]澳斯康生物(南通)股份有限公司,江苏南通221600

出　　处：《中南药学》2023年第3期625-631,共7页Central South Pharmacy

摘　　要：目的采用一种高效的中国仓鼠卵巢细胞(CHO)细胞株构建方法获得稳定高表达治疗性重组蛋白的CHO细胞株。方法利用自主研发载体构建含目的基因的真核表达质粒,经电击转染转入CHO细胞中,经抗性压力筛选后使用细胞池至单克隆的一步筛选策略,配合一种新型的细胞成像系统,高效、快速地筛选出高表达单克隆株。结果成功构建多株稳定的高表达单克隆细胞株,其产量最高可至5.2 g·L^(-1);实验通过毛细管等电聚焦电泳法等进一步验证了细胞株产物关键质量特性,等电点均为9.2,所得抗体等电点较好。结论相比传统的构建方法需6~8个月,本研究筛选得到的高表达单克隆细胞株周期缩短了2个月。Objective To obtain stable Chinese hamster ovary(CHO)cell lines with a high expression of single-type therapeutic recombinant proteins by a high-efficiency cell line development method.Methods Electroporation was used to transfect CHO cells with a self-developed vector to construct the eukaryotic expression plasmid containing the target genes.After the drug resistance screening of the CHO cell pool,a one-step screening strategy was used to rapidly filter the high-expression monoclonal strains,with support from a new cell imaging system.Results Several stable highexpression monoclonal cell lines were successfully constructed,and the highest output reached 5.2 g·L^(-1).The characteristics of the cell line products were further verified via capillary isoelectric focusing electrophoresis.The isoelectric point of clones was 9.2,which was satisfactory.Conclusion In comparison with the 6~8 months by traditional construction method,the entire experiment of the high-expression monoclonal cell lines is shortened by 2 months.

关 键 词：中国仓鼠卵巢细胞 细胞株构建 高效 筛选策略 

分 类 号：R392[医药卫生—免疫学]