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A functional analysis of N-glycosylation-related genes on sialylation of recombinant erythropoietin in six commonly used mammalian cell lines

Paper ID Volume ID Publish Year Pages File Format Full-Text
31705 44831 2010 11 PDF Available
Title
A functional analysis of N-glycosylation-related genes on sialylation of recombinant erythropoietin in six commonly used mammalian cell lines
Abstract

Significant efforts have been made to improve the sialylation of recombinant glycoproteins with the aim of extending their in vivo circulation time. Here, we report a systematic functional analysis of 31 N-glycosylation-related genes on sialylation of recombinant EPO in six cell lines. BHK and CHO cells were found to sialylate recombinant EPO most effectively. None of the 31 genes, individually or in combination, was able to improve EPO sialylation in these cells. HEK293, Cos-7 and 3T3 cells showed intermediate sialylation capabilities, whereas NS0 cells sialylated recombinant EPO poorly. Overexpression of ST6GalI, ST3GalIII or ST3GalIV, but not ST3GalVI, was able to improve EPO sialylation in these four cell lines. qRT-PCR experiments revealed that ST3GalIII and ST3GalIV are indeed under expressed in HEK293, 3T3 and NS0 cells. Co-expression of upstream glycogenes failed to synergize with these sialyltransferases to further enhance sialylation, suggesting that the upstream glycogenes are all expressed at sufficient levels.

Keywords
Glycogenes; Isoelectric focusing (IEF); N-glycosylation; Recombinant erythropoietin (EPO); Sialic acidFuc, fucose; Gal, galactose; Glc, glucose; GlcNAc, N-acetylglucosamine; MAA, Maackia amurensis agglutinin; Man, mannose; Neu5Ac, N-acetylneuramic acid; S
First Page Preview
A functional analysis of N-glycosylation-related genes on sialylation of recombinant erythropoietin in six commonly used mammalian cell lines
Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 12, Issue 6, November 2010, Pages 526–536
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering