文献类型：期刊文献

英文题名：Insights into the loss of protein sialylation in an fc-fusion protein-producing CHO cell bioprocess

作者：Chen, Xinning[1];Liu, Xuping[1];Xiao, Zheng[1];Liu, Jintao[1];Zhao, Liang[1];Tan, Wen-Song[1];Fan, Li[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2019

卷号：103

期号：12

起止页码：4753

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20192006914272);WOS:【SCI-EXPANDED(收录号:WOS:000469192100008)】；

基金：This work was supported by the Major Programs of Development Foundation of Shanghai Zhangjiang National Independent Innovation Demonstration Zone (No. ZJ2015-ZD-002).

语种：英文

外文关键词：Chinese hamster ovary cells; Fc-fusion protein; Sialylation; Extracellular degradation; Intracellular biosynthesis

摘要：Sialylation affects circulating half-life, charge distribution, and other biochemical properties of therapeutic glycoproteins. Loss of protein sialylation during glycoprotein-producing bioprocesses could lead to a low final protein sialylation level and bring negative effects on subsequent clinical efficacy. In this work, an Fc-fusion protein-producing Chinese hamster ovary cell fed-batch culture process was studied and insights into the loss of protein sialylation during the Fc-fusion protein production phase (days 5 to 13) were presented. The results showed that the decreased total sialic acid content was 13.84 g/mg during the production phase, which accounted for 24% of the total sialic acid content on day 5. The lost sialic acids were predominantly from 2-3 sialylation on N- and O-glycans. Through cell-free incubation and kinetics studies, it was found that the decreased sialic acid content caused by extracellular sialic acid degradation and incomplete glycan biosynthesis were 7.79 g/mg and 6.05 g/mg, respectively. The two processes had a nearly equal contribution to the loss of final product sialylation. Detailed characterizations revealed that decreases in sialic acid content were due either to extracellular sialic acid degradation via hydrolysis of 2-3 sialic acids probably by released cytosolic sialidase or to a lack of galactosylated glycan availability for sialylation during late-stage glycosylation. Our work provides a better understanding of losses in protein sialylation during glycoprotein manufacturing.