恒稳磁场辅助CHO细胞培养和表达特性研究  

Cultivation and Expression of Monoclonal Chinese Hamster Ovary Cells Under Constant Magnetic Field

作　　者：李晓红 李红文 高丽 张雪艳 杨哪[2] 李荣明 LI Xiaohong;LI Hongwen;GAO Li;ZHANG Xueyan;YANG Na;LI Rongming(Process Development,RemeGen Co.,Ltd.,Yantai 264000,China;School of Food Science and Technology,Jiangnan University,Wuxi 214122,China;R&D Department,INDUC Scientific Co.,Ltd.,Wuxi 214153,China)

机构地区：[1]荣昌生物制药(烟台)股份有限公司,山东烟台264006 [2]江南大学食品学院,江苏无锡214122 [3]英都斯特(无锡)感应科技有限公司,江苏无锡214025

出　　处：《工业微生物》2024年第5期39-43,共5页Industrial Microbiology

基　　金：国家自然科学基金面上项目(编号:32172353);江苏省优秀青年基金项目(编号:BK20211582);无锡市科技发展资金项目(编号:G20223003)。

摘　　要：以中国仓鼠卵巢细胞为宿主细胞构建重组表达细胞用以表达单克隆抗体,已经被广泛应用于生物制药领域。物理调控是影响细胞增殖和表达的重要方式,但针对恒稳磁场暴露下的CHO细胞培养及其抗体表达还鲜有报道。基于此,文章利用3.0 mT恒稳磁场,在37℃、8%CO_(2)环境下对CHO细胞进行克隆培养,通过克隆扩增筛选、蛋白表达及分析、拷贝数监测等手段来评估其对抗体表达的影响。结果表明,与对照组相比,在3.0 mT磁场环境下,单克隆形成数量增加了13.24%,细胞数量显著增多,单克隆抗体表达水平提升53.2%,克隆筛选周期缩短4~5 d。此外,在亚克隆筛选中,亚克隆的细胞数量和抗体表达在磁场暴露下略有增加,而抗体表达、蛋白质量和基因拷贝数在传代后均无显著变化,表明磁场暴露环境中培养的重组CHO细胞具有稳定的性质。该研究结果有望为未来CHO细胞的大规模培养以及磁场暴露下的高效生产抗体研究提供参考。The use of Chinese hamster ovary cells as host cells to construct recombinant expression cells for the expression of monoclonal antibodies has been widely used in the field of biopharmaceuticals.Physical regulation is an important way to influence the cell proliferation and expression,and there are few reports on CHO cell culture and antibody expression under constant constant magnetic field exposure.In view of this,in this study,CHO cells were clonally cultured using a constant magnetic field of 3.0 mT at 37益and 8%CO_(2),and the effect of the constant magnetic field on antibody expression were evaluated by clonal expansion screening,protein expression and analysis,and copy number monitoring.The results showed that compared with the control,the number of monoclonal formation was increased by 13.24%,the cell number was significantly increased,the antibody expression level of monoclonal was elevated by 53.2%,and the clone screening cycle was shortened by 4-5 days under the 3.0 mT magnetic field environment.In addition,in the subclonal screening,the cell number and antibody expression of subclones were slightly increased under the magnetic field exposure,and there was no significant change in antibody expression,protein amount and gene copy number after passaging,indicating that the nature of recombinant CHO cells cultured by magnetic field exposure was stable.The results of this study can provide references for future researches on large-scale cultivation of CHO cells as well as the efficient production of antibodies under magnetic field exposure.

关 键 词：CHO细胞 恒稳磁场 克隆表达 抗体表达 蛋白质特性 

分 类 号：Q813.11[生物学—生物工程]