文献类型：期刊文献

英文题名：Galactose supplementation enhance sialylation of recombinant Fc-fusion protein in CHO cell: an insight into the role of galactosylation in sialylation

作者：Liu, Jintao[1];Wang, Jie[1];Fan, Li[1];Chen, Xinning[1];Hu, Dongdong[1];Deng, Xiancun[2];Poon, H. Fai[2];Wang, Haibin[2];Liu, Xuping[1];Tan, Wen-Song[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Zhejiang Hisun Pharmaceut Hangzhou Co Ltd, Hangzhou 311404, Zhejiang, Peoples R China

年份：2015

卷号：31

期号：7

起止页码：1147

外文期刊名：WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY

收录：;EI(收录号：20151800812828);WOS:【SCI-EXPANDED(收录号:WOS:000355728400014)】；

基金：This work was supported by the National Natural Science Foundation of China (No. 21206040, 21406066), the National High Technology Research and Development Program of China (863 Program) (No. 2012AA02A303), the National Science and Technology Major Project (No. 2013ZX10004003-003-003), the Fundamental Research Funds for the Central Universities (WF1214035).

语种：英文

外文关键词：Fc-fusion protein; CHO cell; Glycosylation; dSialylation; Galactose; Galactosylation

摘要：Sialic acid levels of therapeutic glycoprotein play an important role in plasma half-life. An undesirable decrease of sialic acid content was observed when we increased Fc-fusion protein productivity fourfold in a GS-CHO cell line by bioprocess optimization. We investigated the potential mechanism for the sialic acid content reduction. We found that limited nucleotide sugar precursor and the extracellular sialidase were not responsible for the reduction of the sialic acid content after titer improvement. Oligosaccharide analysis revealed that the lack of protein galactosylation was the potential cause for the reduction of sialic acid content. Thus we validated this notion by evaluated galactose supplementation in 2 L bioreactors. Cell culture performance was not impacted by addition of up to 40 mM galactose except for the glucose consumption rate. Addition of 20 mM galactose to the bioreactor resulted in the increase of 44 % for total sialic acid content and 20.3 % for sialylated glycans. These data were further validated when the process was run on 200 L scaled bioreactor. These data together show that the galactosylation plays an apparent role in sialylation in our current system.