Digital mRNA profiling of N-glycosylation gene expression in recombinant Chinese hamster ovary cells treated with sodium butyrate

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To understand the effects of sodium butyrate (NaBu) on protein glycosylation, recombinant Chinese hamster ovary (rCHO) cells producing Fc-fusion glycoprotein were subjected to 3 mM NaBu. The addition of NaBu to the cultures reduced the relative proportion of acidic isoforms and sialic acid content of the glycoprotein. Fifty-two N-glycosylation-related gene expressions were also assessed by the NanoString nCounter system, which can provide a direct digital readout using custom-designed color-coded probes. Among them, ten genes (ugp, slc35a2, ganc, man1a, man1c, mgat5a, st3gal5, glb1, neu1, and neu3) were up-regulated and three genes (b4galt2, st3gal3, and neu2) were down-regulated significantly. Altered expression patterns in st3gal3, neu1, and neu3, which have roles in the sialic acid biosynthesis pathway, correlated with reduced sialic acid content of the glycoprotein by NaBu. Taken together, the results obtained in this study provide a better understanding of the detrimental effect of NaBu on N-glycosylation in rCHO cells.
Publisher
ELSEVIER SCIENCE BV
Issue Date
2014-02
Language
English
Article Type
Article
Keywords

LOW CULTURE TEMPERATURE; CHO-CELLS; MAMMALIAN-CELLS; ERYTHROPOIETIN; TRANSCRIPTOME; PRODUCTIVITY; SIALYLATION; HETEROGENEITY; PROTEIN; BATCH

Citation

JOURNAL OF BIOTECHNOLOGY, v.171, no.1, pp.56 - 60

ISSN
0168-1656
DOI
10.1016/j.jbiotec.2013.12.001
URI
http://hdl.handle.net/10203/189578
Appears in Collection
BS-Journal Papers(저널논문)
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