CHO细胞表达糖蛋白的研究进展<sup>*</sup>

doi:10.13523/j.cb.2201001

中国生物工程杂志

2022, Vol. 42

Issue (6): 54-65 DOI: 10.13523/j.cb.2201001

综述

CHO细胞表达糖蛋白的研究进展^*

任自强^1,²,王梦灿^1,²,张海灵¹,朱希强^2,^**(

),林剑^1,^**(

)

1.烟台大学生命科学学院烟台 264005
2.山东丰金生物医药有限公司烟台 264117

Research Progress of Glycoprotein Expression in CHO Cells

REN Zi-qiang^1,²,WANG Meng-can^1,²,ZHANG Hai-ling¹,ZHU Xi-qiang^2,^**(

),LIN Jian^1,^**(

)

1. College of Life Sciences, Yantai University, Yantai 264005, China
2. Shandong Fengjin Biomedical Co., Ltd., Yantai 264117, China

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摘要：

中国仓鼠卵巢细胞(Chinese hamster ovary cells,CHO)表达系统因具有较高密度培养、高表达和相对完整的蛋白质糖基化修饰系统等特点,成为生产糖蛋白广泛应用的宿主表达细胞之一。目前已产生不同的CHO细胞系和各种功能细胞株以满足对糖蛋白的大量生产和其他实验需求。近年来,随着基因工程、蛋白质工程、细胞工程和发酵调控等技术的发展应用,由CHO细胞生产糖蛋白的产量和糖基化修饰程度取得了突破。然而,随着生物制品市场对于糖蛋白的需求增加,如何获得大量、均质的糖蛋白也成为急需解决的问题。综述了不同工程CHO表达系统的研究、应用、糖基化修饰系统,以及影响外源糖蛋白在CHO系统表达和糖基化修饰的理化因素,结合文献总结并预测了未来CHO细胞表达系统研究的四个具有重大意义的研究方向,以期在未来可以改善由CHO细胞表达糖蛋白的产量和质量。

关键词： 中国仓鼠卵巢细胞; 糖蛋白表达; 糖基化

Abstract:

The Chinese hamster ovary cells(CHO) cell expression system has become a widely used host expression system for the production of glycoproteins due to its high-density culture, high expression characteristics and relatively complete protein glycosylation modification system. Different CHO cell lines and various functional cell lines have been generated to date to meet the needs of large-scale production of glycoproteins and other experimental purposes. In recent years, with the development and application of genetic engineering, protein engineering, cell engineering, fermentation regulation and other technologies, breakthroughs have been made in the yield and degree of glycosylation of glycoproteins produced by CHO cells. However, with the increasing demand for glycoproteins in the biological product market, how to obtain a large quantity of glycoproteins with uniform mixture has also become an urgent problem to be solved. This review introduces the application of CHO and the research progress on gene construction and expression, enzyme engineering, cell lines, molecular chaperones, additives and physical conditions that affect the yield of expressed exogenous proteins and the degree of glycosylation modification in CHO cells. Combined with literature analysis, four directions for future CHO cell research are predicted: the development of new engineered CHO cell lines, a stable CHO expression system, synergistic strategies, and multi-omics applications. It is expected that the yield and quality of glycoproteins expressed by CHO cells can be improved in the future to meet the needs of clinical studies and research.

Key words: Chinese hamster ovary cells(CHO) Glycoprotein expression Glycosylation

收稿日期: 2022-01-05 出版日期: 2022-07-07

ZTFLH:

Q786

基金资助: *国家自然科学基金(32001836);山东省自然科学基金(ZR2020QC233)

通讯作者: 朱希强,林剑 E-mail: 15066696818@163.com;linjian3384@163.com

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引用本文:

任自强,王梦灿,张海灵,朱希强,林剑. CHO细胞表达糖蛋白的研究进展^*[J]. 中国生物工程杂志, 2022, 42(6): 54-65.

REN Zi-qiang,WANG Meng-can,ZHANG Hai-ling,ZHU Xi-qiang,LIN Jian. Research Progress of Glycoprotein Expression in CHO Cells. China Biotechnology, 2022, 42(6): 54-65.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2201001 或 https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I6/54

表1 CHO 细胞产生的糖蛋白列表

表2 上调或敲除基因对CHO细胞系表达时间的影响

图1 CHO细胞中蛋白质的天冬酰胺糖基化代谢合成途径

表3 糖基化工程细胞的表达和结果

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