HEK293 PEAK瞬转快速表达抗体蛋白平台的建立

The platform of transient transfection for antibody expressed in HEK293 PEAK cells

将HEK293 PEAK细胞用Gibco FreeStyleTM293表达培养液进行无血清悬浮驯化培养,获得无血清悬浮培养的HEK293 PEAK细胞,并经有限稀释法获得细胞亚克隆,建立抗体瞬转表达的工程细胞株.用转染试剂PEI将质粒转入该细胞株中表达抗体.通过优化质粒DNA与转染试剂PEI的比例,质粒与转染试剂的比例为1∶2时,转染效率最佳,并对瞬转表达抗体进行鉴定和活性分析,确认获得具有生物活性的抗体(Herceptin),抗体分子量约150 kD,结果表明已成功建立了以无血清悬浮培养的HEK293 PEAK细胞为宿主细胞的瞬转快速表达抗体蛋白平台.

This research established engineering cell line for transient transfection for antibody by adapting with Gibco （@） FreeStyleTM 293 expression medium and limited dilution.The ratio of antibody plasmid and PEI （transfection reagent） was optimized to be that the ratio of 1∶2 （antibody plasmid∶ PEI） is the best result.The molecular weight of antibody expressed in HEK293 PEAK cells is about 150 kD,and the biological activity was similar to control antibody（Herceptin）.In summary we successfully established the platform for rapidly expressing antibody in HEK293 PEAK cells cultured in serum-free medium.