文献类型：期刊文献

英文题名：Enhancement of Recombinant Human EPO Production and Sialylation in Chinese Hamster Ovary Cells Through Bombyx mori 30Kc19 Gene Expression

作者：Wang, Zesong[1,2];Park, Ju Hyun[1];Park, Hee Ho[1];Tan, Wensong[2];Park, Tai Hyun[1]

机构：[1]Seoul Natl Univ, BioMAX Inst, Sch Chem & Biol Engn, Seoul 151744, South Korea;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2011

卷号：108

期号：7

起止页码：1634

外文期刊名：BIOTECHNOLOGY AND BIOENGINEERING

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000291467600015)】；

基金：Contract grant sponsor: National Research Foundation of Korea (NRF) Contract grant number: 2010K001137; 2010-0000825; 20100020821; R32-2010-000-10213-0 This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (No. 2010K001137, 2010-0000825, 20100020821). This work was also supported by WCU (World Class University) program through the Korea Science and Engineering Foundation funded by the Ministry of Education, Science and Technology (R32-2010-000-10213-0).

语种：英文

外文关键词：CHO cell; EPO; 30Kc19; productivity; sialylation; mitochondrial membrane potential; ATP; sialyltransferase

摘要：Productivity and sialylation are two important factors for the production of recombinant glycoproteins in mammalian cell culture. In our previous study, we found that silkworm hemolymph increased the sialylation of recombinant secreted human placental alkaline phosphatase in the insect cells, promoted the transfer of sialic acids onto the glycoprotein oligosaccharides in an in vitro asialofetuin sialylation system, and enhanced recombinant protein production in the Chinese hamster ovary (CHO) cells. These beneficial effects were mainly due to the 30K proteins, which consist of five isoforms. Among the 30K proteins, 30Kc19 was determined to be the major component. In this study, the 30Kc19 gene was introduced into a CHO cell line producing recombinant human erythropoietin, and its effects on productivity and sialylation were investigated. The transient expression of 30Kc19 significantly improved the production and sialylation of EPO. A stable cell line containing 30Kc19 was also established to investigate the effect of 30Kc19 gene expression. The stable expression of 30Kc19 increased the production and sialylation by 102.6% and 87.1%, respectively. The enhanced productivity from 30Kc19 expression is believed to occur because the 30Kc19 protein suppresses the loss of mitochondrial membrane potential and consequently improves the generation of intracellular ATP. In addition, the positive effect of 30Kc19 expression on sialylation is believed to be due to its ability to maintain sialyltransferase activity. In conclusion, 30Kc19 expression is a novel approach to improve the production and sialylation of recombinant glycoproteins in CHO cells. Biotechnol. Bioeng. 2011; 108: 1634-1642. (C) 2011 Wiley Periodicals, Inc.