A novel sugar analog enhances sialic acid production and biotherapeutic sialylation in CHO cells

Bojiao Yin, Qiong Wang, Cheng Yu Chung, Rahul Bhattacharya, Xiaozhi Ren, Juechun Tang, Kevin J. Yarema, Michael J. Betenbaugh

Research output: Contribution to journalArticlepeer-review

Abstract

A desirable feature of many therapeutic glycoprotein production processes is to maximize the final sialic acid content. In this study, the effect of applying a novel chemical analog of the sialic acid precursor N-acetylmannosamine (ManNAc) on the sialic acid content of cellular proteins and a model recombinant glycoprotein, erythropoietin (EPO), was investigated in CHO-K1 cells. By introducing the 1,3,4-O-Bu3ManNAc analog at 200–300 µM into cell culture media, the intracellular sialic acid content of EPO-expressing cells increased ∼8-fold over untreated controls while the level of cellular sialylated glycoconjugates increased significantly as well. For example, addition of 200–300 µM 1,3,4-O-Bu3ManNAc resulted in >40% increase in final sialic acid content of recombinant EPO, while natural ManNAc at ∼100 times higher concentration of 20 mM produced a less profound change in EPO sialylation. Collectively, these results indicate that butyrate-derivatization of ManNAc improves the capacity of cells to incorporate exogenous ManNAc into the sialic acid biosynthetic pathway and thereby increase sialylation of recombinant EPO and other glycoproteins. This study establishes 1,3,4-O-Bu3ManNAc as a novel chemical supplement to improve glycoprotein quality and sialylation levels at concentrations orders of magnitude lower than alternative approaches. Biotechnol. Bioeng. 2017;114: 1899–1902.

Original languageEnglish (US)
Pages (from-to)1899-1902
Number of pages4
JournalBiotechnology and bioengineering
Volume114
Issue number8
DOIs
StatePublished - Aug 2017

Keywords

  • Chinese hamster ovary (CHO) media
  • N-acetylmannosamine (ManNAc) analog
  • glycosylation
  • metabolic oligosaccharide engineering
  • sialylation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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