CHO细胞中通过基因过表达提高治疗性蛋白产量的策略

The strategy for enhancing therapeutic protein yield by gene overexpression in CHO cells

中国仓鼠卵巢(Chinese hamster ovary,CHO)细胞是重要的表达治疗性糖蛋白的工程细胞。CHO细胞能够生产高质量的生物制剂,与人类拥有相似的翻译后修饰功能。与细菌和酵母表达系统相比,CHO表达系统还存在细胞生长明显受限、产量相对低、抗压能力弱、成本昂贵等缺点。生产工艺优化已经是CHO细胞中治疗性重组蛋白表达量大幅提升的重要手段,此外,对特定基因进行过表达、敲除或者转录后沉默等基因工程方法也为CHO细胞工程提供了强有力的理论和实践基础。较全面地综述了近20年过表达手段与CHO细胞工程改造之间的关系,并以细胞的增殖和死亡、代谢和分泌、转录后翻译以及对蛋白的糖基化修饰为主要研究方向,讨论了最新过表达策略的研究进展和应用潜力。

Chinese hamster ovary(CHO)cells are important engineering cells expressing therapeutic glycoprotein.CHO cells are capable of producing high quality biologicals with similar post-translational modifications to humans.Although biological process optimization has greatly increased the expression of therapeutic recombinant proteins in CHO cells,there are still many constraints.Compared with bacterial and yeast expression systems,the CHO expression system is characterized by markedly restricted cell growth,relatively low yield,weak stress resistance and high cost.In addition to optimizing production processes,gene editing such as overexpression,knockout or post-transcriptional silencing of specific genes has provided a strong theoretical and practical basis for CHO cell engineering.This review comprehensively summarizes the relationship between overexpression strategy and CHO cells engineering during the past 20 years and discusses the latest research progress and potential application of overexpression strategies targeting cell proliferation and death,metabolism and secretion,post-transcription translation,and glycosylation of proteins.