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Abstract：
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.
摘要：
昆虫细胞最近被证明是高水平生产功能性重组蛋白的极好平台。自噬是通过消除受损的细胞器和蛋白质聚集体来促进细胞存活的重要机制。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,和氯化锂作用于分泌工程化抗体分子的重组鳞翅目昆虫细胞。与不治疗相比，用雷帕霉素或依维莫司治疗延长细胞生长，以允许高细胞密度，在下降阶段提高生存能力，然后增加分泌抗体的产量。这些积极作用似乎是通过自噬诱导的，因为自噬体被清楚地检测到，特别是在用雷帕霉素或依维莫司处理的细胞中。不像雷帕霉素,另一种自噬诱导剂，FK506在昆虫细胞中无效。添加适当的自噬诱导剂可有效提高重组蛋白在昆虫细胞中的生产率。
