You Min, Liu Yanning, Chen Yingwei, Guo Jiyuan, Wu Jun, Fu Yajuan, Shen Rong, Qi Rui, Luo Wenxin, Xia Ningshao
National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, China.
Biosci Biotechnol Biochem. 2013;77(6):1207-13. doi: 10.1271/bbb.120968. Epub 2013 Jun 7.
Due to the great diversity in protein expression productivity, a customized transient gene expression (TGE) method was used in the present study to optimize transient expression of three antibodies. Several factors, including host cells, temperature, valproic acid (VPA) treatment, various vectors, and additives were optimized independently and then combined to form a customized TGE protocol for each antibody. In the event, the optimized TGE conditions for three antibodies were different from each other. Compared with the TGE in CHO-S cells by pCDNA3.1 expression vector, the expression productivities of 8C11 cAb, 37 hAb, and 10F7 cAb showed 16-fold, 293-fold, and 19-fold increases respectively by the customized TGE method. For 8C11 cAb, coexpressing L-chain and H-chain on different plasmids led to higher yields. The customized TGE method is an alternative approach that can greatly improve the expression productivity of a variety of recombinant proteins.
由于蛋白质表达生产力存在巨大差异，本研究采用定制的瞬时基因表达（TGE）方法来优化三种抗体的瞬时表达。包括宿主细胞、温度、丙戊酸（VPA）处理、各种载体和添加剂在内的几个因素被分别优化，然后组合形成针对每种抗体的定制TGE方案。结果，三种抗体的优化TGE条件各不相同。与使用pCDNA3.1表达载体在CHO-S细胞中进行的TGE相比，定制的TGE方法使8C11单克隆抗体（cAb）、37人源化抗体（hAb）和10F7 cAb的表达生产力分别提高了16倍、293倍和19倍。对于8C11 cAb，在不同质粒上共表达轻链和重链可提高产量。定制的TGE方法是一种替代方法，可大大提高多种重组蛋白的表达生产力。
