Nakanuma Ryou, Masumi-Koizumi Kyoko, Ohmuro-Matsuyama Yuki, Katsuda Tomohisa, Yamaji Hideki
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
Cytotechnology. 2020 Sep 29;73(3):299-305. doi: 10.1007/s10616-020-00423-6.
Insect cells have recently proven to be an excellent platform for the high-level production of functional recombinant proteins. Autophagy is an important mechanism that promotes cell survival by eliminating damaged organelles and protein aggregates, and it also may influence recombinant protein production. In the present study, we compared the effects that autophagy inducers rapamycin, everolimus, and lithium chloride exert on recombinant lepidopteran insect cells that secrete an engineered antibody molecule. Compared with nontreatment, treatment with either rapamycin or everolimus prolonged cell growth to allow high cell density, improved viability in the declining phase, and then increased the yield of secreted antibodies. These positive effects appeared to be induced via autophagy since autophagosomes were clearly detected, particularly in cells treated with rapamycin or everolimus. Unlike rapamycin, another autophagy inducer, FK506, was ineffective in insect cells. The addition of an appropriate autophagy inducer may be effective in increasing the productivity of recombinant proteins in insect cells.
昆虫细胞最近已被证明是用于高水平生产功能性重组蛋白的优良平台。自噬是一种重要机制,通过清除受损细胞器和蛋白质聚集体来促进细胞存活,并且它也可能影响重组蛋白的生产。在本研究中,我们比较了自噬诱导剂雷帕霉素、依维莫司和氯化锂对分泌工程化抗体分子的重组鳞翅目昆虫细胞的影响。与未处理相比,用雷帕霉素或依维莫司处理可延长细胞生长以实现高细胞密度,改善衰退期的活力,进而提高分泌抗体的产量。这些积极作用似乎是通过自噬诱导的,因为清楚地检测到了自噬体,特别是在用雷帕霉素或依维莫司处理的细胞中。与雷帕霉素不同,另一种自噬诱导剂FK506在昆虫细胞中无效。添加合适的自噬诱导剂可能有效地提高昆虫细胞中重组蛋白的生产力。