抗人PD-L1单克隆抗体的瞬转表达

吉林大学学报(理学版)

抗人PD-L1单克隆抗体的瞬转表达

史晶莹^1, 郭晶^1, 李雪^1, 董雪^1, 孙博^2, 吴丛梅^1, 殷玉和¹

1. 长春工业大学化学与生命科学学院, 长春 130012； 2. 吉林大学生命科学学院, 长春 130012

收稿日期:2017-08-15 出版日期:2018-07-26 发布日期:2018-07-31
通讯作者: 殷玉和 E-mail:yyhlxt72@163.com

Transient Expression of Antihuman PDL1 Monoclonal Antibodies

SHI Jingying¹, GUO Jing¹, LI Xue^1, DONG Xue^1, SUN Bo^2, WU Congmei^1, YIN Yuhe^1

1. College of Chemistry and Life Science, Changchun University of Technology, Changchun 130012, China;2. College of Life Sciences, Jilin University, Changchun 130012, China

Received:2017-08-15 Online:2018-07-26 Published:2018-07-31
Contact: YIN Yuhe E-mail:yyhlxt72@163.com

摘要/Abstract

摘要： 利用HEK293 6E细胞瞬转表达抗人PDL1单克隆抗体. 将HEK293 6E细胞无血清悬浮培养, 筛选最佳生长培养基; 考察HEK293 6E细胞的筛选表达培养基、 DNA和PEI质量比及转染时细胞密度等转染条件, 并对收获的抗体进行SDSPAGE和ELISA鉴定. 结果表明: 在A无血清表达培养基中, 以2×106个/mL的细胞密度和m(DNA)∶m(PEI)=1的条件转染, 第4天收获抗体的产量最高, 为170.9 μg/mL; 收获PD-L1抗体的重链分子量约为50 000, 轻链分子量约为25 000.

关键词: 转染, 抗人PD-L1, 抗体, 瞬转表达

Abstract: The PDL1 monoclonal antibody was transiently expressed by using HEK293 6E cells. We cultivated HEK293 6E cells in serumfree suspension and selected the optimum growth medium, investigated the transfection conditions of HEK293 6E cells, including culture medium varieties, the mass ratio of DNA and PEI, and cell density while transfecting, then identified the harvested antibodies by SDSPAGE and ELISA. The results show that the best transfection conditions were as follows: A style, a serumfree culture medium, a cell density of 2×106 cells/mL and m(DNA)∶m(PEI)=1. The highest yield of antibody was 170.9 μg/mL at the fourth day after transfection, the molecular weight of the heavy chain of the harvested PDL1 antibody was about 50 000 and that of the light chain was about 25 000.

Key words: anti-human PD-L1, transfection, transient expression, antibody

中图分类号:

史晶莹, 郭晶, 李雪, 董雪, 孙博, 吴丛梅, 殷玉和. 抗人PD-L1单克隆抗体的瞬转表达[J]. 吉林大学学报(理学版), 2018, 56(4): 1024-1028.

SHI Jingying, GUO Jing, LI Xue, DONG Xue, SUN Bo, WU Congmei, YIN Yuhe. Transient Expression of Antihuman PDL1 Monoclonal Antibodies[J]. Journal of Jilin University Science Edition, 2018, 56(4): 1024-1028.

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