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

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

作者机构：Chinese Acad Sci Shenzhen Inst Synthet Biol Shenzhen Inst Adv Technol Key Lab Quantitat Synthet Biol Shenzhen 518055 Peoples R China Shenzhen Weiguang Biol Prod Co Ltd Shenzhen 518107 Peoples R China Shenzhen Innovat Pharmaceut Engn Lab Recombinant P Shenzhen 518107 Peoples R China 

出 版 物：《PROTEIN EXPRESSION AND PURIFICATION》 (Protein Expr. Purif.)

年 卷 期：2025年第229卷

页      面：106666页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 07[理学] 0836[工学-生物工程] 

基　　金：National Key R&D Program of China [2020YFA0908800] Shenzhen Science and Technology Innovation Committee of China [JSGG20200207155251653] 

主　　题：Blood coagulation factor VII PiggyBac transposon system CHO cells Recombinant protein Production Fed-batch 

摘      要：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.