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

Paper ID Volume ID Publish Year Pages File Format Full-Text
23250 43424 2014 5 PDF Available
Title
Digital mRNA profiling of N-glycosylation gene expression in recombinant Chinese hamster ovary cells treated with sodium butyrate
Abstract

•Recombinant CHO (rCHO) cells are subjected to sodium butyrate (NaBu).•N-Glycosylation-related gene expressions are assessed by NanoString nCounter system.•Altered gene expressions are correlated with sialic acid content of the glycoprotein.•The results provide a better understanding of the effect of NaBu on N-glycosylation.

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.

Keywords
Sodium butyrate; N-Glycosylation; Gene expression; NanoString nCounter system; CHO cells
First Page Preview
Digital mRNA profiling of N-glycosylation gene expression in recombinant Chinese hamster ovary cells treated with sodium butyrate
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 171, 10 February 2014, Pages 56–60
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering