Efficient production of recombinant human FVII in CHO cells using the piggyBac transposon system

Zhen Yang; Xueyun Wang; Shan Luo; Hui Li; Jiangbo Xu; Linlin Liang; Zhimin He; Guangyuan Wang; Zhuobin Wu; Nan Zhong; Haijun Xiang; Zhan Zhang; Caiping Guo; Yunjia Zhang; Fei Yan

doi:10.1016/j.pep.2025.106666

Efficient production of recombinant human FVII in CHO cells using the piggyBac transposon system

Protein Expr Purif. 2025 Jan 21:106666. doi: 10.1016/j.pep.2025.106666. Online ahead of print.

Authors

Zhen Yang¹, Xueyun Wang², Shan Luo², Hui Li², Jiangbo Xu², Linlin Liang², Zhimin He², Guangyuan Wang², Zhuobin Wu², Nan Zhong², Haijun Xiang², Zhan Zhang², Caiping Guo³, Yunjia Zhang⁴, Fei Yan⁵

Affiliations

¹ Paul C. Lauterbur Research Center for Biomedical Imaging, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, P.R. China, 518055; Shenzhen Weiguang Biological Products Co., Ltd., Shenzhen, P.R. China, 518107; Shenzhen Innovation Pharmaceutical Engineering Laboratory for Recombinant Plasma Proteins, Shenzhen 518107, China.
² Shenzhen Weiguang Biological Products Co., Ltd., Shenzhen, P.R. China, 518107; Shenzhen Innovation Pharmaceutical Engineering Laboratory for Recombinant Plasma Proteins, Shenzhen 518107, China.
³ Shenzhen Weiguang Biological Products Co., Ltd., Shenzhen, P.R. China, 518107; Shenzhen Innovation Pharmaceutical Engineering Laboratory for Recombinant Plasma Proteins, Shenzhen 518107, China. Electronic address: gcpzxy0402@aliyun.com.
⁴ Shenzhen Weiguang Biological Products Co., Ltd., Shenzhen, P.R. China, 518107; Shenzhen Innovation Pharmaceutical Engineering Laboratory for Recombinant Plasma Proteins, Shenzhen 518107, China. Electronic address: zhangyunjia@aliyun.com.
⁵ Paul C. Lauterbur Research Center for Biomedical Imaging, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, P.R. China, 518055; Shenzhen Weiguang Biological Products Co., Ltd., Shenzhen, P.R. China, 518107. Electronic address: fei.yan@siat.ac.cn.

PMID: 39848303
DOI: 10.1016/j.pep.2025.106666

Abstract

As an important coagulation factor, activated coagulation factor VII (FVIIa) is mainly used to treat the bleeding of hemophilia patients who have developed inhibitory antibodies against FVIII and FIX conventional treatment. Recombinant human factor VII (rhFVII) produced in mammalian cell lines have been developed as the most important resource of FVIIa. However, cell lines express rhFVII protein derived from an exogenous expression vector at a lower level than most other proteins. In the current study, we have shown efficient rhFVII production in CHO cell lines using piggyBac (PB) transposon system. rhFVII is successfully expressed in fed-batch culture of CHO cells, and the expression of rhFVII up to 100 mg/L. Moreover, the purified secreted rhFVII was determined by SDS-PAGE and Western Blot. The coagulation activity was determined by the chromogenic Activity ELISA kit. In conclusion, this study has demonstrated that the piggyBac transposon system can be used for an efficient production of recombinant FVII.

Keywords: Blood coagulation factor VII; CHO cells; Fed-batch; PiggyBac transposon system; recombinant protein production.