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Applied Microbiology and Biotechnology

, Volume 103, Issue 12, pp 4753–4765 | Cite as

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

  • Xinning Chen
  • Xuping Liu
  • Zheng Xiao
  • Jintao Liu
  • Liang Zhao
  • Wen-Song TanEmail author
  • Li FanEmail author
Biotechnological products and process engineering
  • 229 Downloads

Abstract

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 μg/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 α 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 μg/mg and 6.05 μg/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 α 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.

Keywords

Chinese hamster ovary cells Fc-fusion protein Sialylation Extracellular degradation Intracellular biosynthesis 

Notes

Compliance with ethical standards

Funding

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

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_9850_MOESM1_ESM.pdf (310 kb)
ESM 1 (PDF 309 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The State Key Laboratory of Bioreactor EngineeringEast China University of Science and TechnologyShanghaiChina

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