sTNFRII-gAD-Fc融合蛋白在CHO细胞中的高效表达精选.pdfVIP

Immunology Studies 免疫学研究, 2015, 3, 1-11 Published Online February 2015 in Hans. /journal/is /10.12677/is.2015.31001 Efficient Expression of sTNFRII-gAD-Fc Fusion Protein in CHO Cell 1* 2* 1 1 1 1# Caihong Wang , Ying Liu , Qinzhen Cai , Hongmei Yang , Lisha Ma , Jimin Gao 1 Zhejiang Provincial Key Laboratory for Technology Application of Model Organisms, School of Laboratory Medicine, School of Life Science, Wenzhou Medical University, Wenzhou Zhejiang 2 Yantai Central Hospital of Longkou Mining, Yantai Shandong # Email: wch995041@163.com, jimingao64@163.com th rd th Received: May 7 , 2015; accepted: May 23 , 2015; published: May 27 , 2015 Copyright © 2015 by authors and Hans Publishers Inc. This work is licensed under the Creative Commons Attribution International License (CC BY). /licenses/by/4.0/ Abstract In order to produce the soluble TNF receptor (sTNFR) II with good neutralizing activity against TNFα, we constructed the fusion gene sTNFRII-gAD-Fc, which encoded human sTNFRII, the globu- lar domain of adiponectin (gAD) and IgG1 Fc, and efficiently expressed it through a “GC-rich” me- thod for mammalian gene expression in CHO-S cells, and further established the process of se- rum-free suspension fed-batch culture. The fusion gene of sTNFRII-gAD-Fc was obtained by re- combinant PCR, and then cloned into pMH3 expression plasmid with GC-rich motif. The plasmid was electrotransfected into CHO-S cells, which were selected by G418. sTNFR