Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, 565-0871, Japan.
Cytotechnology. 2013 Dec;65(6):993-1002. doi: 10.1007/s10616-013-9631-x. Epub 2013 Sep 13.
To improve antibody production in Chinese hamster ovary (CHO) cells, the humanized antibody-producing CHO DP-12-SF cell line was transfected with the gene encoding activating transcription factor 4 (ATF4), a central factor in the unfolded protein response. Overexpression of ATF4 significantly enhanced the production of antibody in the CHO DP-12-SF cell line. The specific IgG production rate of in the ATF4-overexpressing CHO-ATF4-16 cells was approximately 2.4 times that of the parental host cell line. Clone CHO-ATF4-16 did not show any change in growth rate compared with the parental cells or mock-transfected CHO-DP12-SF cells. The expression levels of mRNAs encoding both the antibody heavy and light chains in the CHO-ATF4-16 clone were analyzed. This analysis showed that ATF4 overexpression improved the total production and specific production rate of antibody without affecting the mRNA transcription level. These results indicate that ATF4 overexpression is a promising method for improving recombinant IgG production in CHO cells.
为了提高中国仓鼠卵巢（CHO）细胞中的抗体产量，将编码激活转录因子 4（ATF4）的基因转染到产生人源化抗体的 CHO DP-12-SF 细胞系中，ATF4 是未折叠蛋白反应的中心因子。ATF4 的过表达显著提高了 CHO DP-12-SF 细胞系中抗体的产生。过表达 ATF4 的 CHO-ATF4-16 细胞的特异性 IgG 产生速率约为亲本宿主细胞系的 2.4 倍。与亲本细胞或 mock 转染的 CHO-DP12-SF 细胞相比，CHO-ATF4-16 克隆的生长速率没有任何变化。分析了 CHO-ATF4-16 克隆中编码抗体重链和轻链的 mRNA 的表达水平。该分析表明，ATF4 的过表达提高了抗体的总产量和特异性产生速率，而不影响 mRNA 转录水平。这些结果表明，ATF4 的过表达是提高 CHO 细胞中重组 IgG 产量的一种很有前途的方法。
