Altering the central carbon metabolism of HEK293 cells: impact on recombinant glycoprotein quality

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DOIResolve DOI: http://doi.org/10.1016/j.jbiotec.2016.12.003
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TypeArticle
Journal titleJournal of Biotechnology
ISSN0168-1656
Volume242
Pages7382
Subjectprotein quality; HEK293 cells; PYC2-overexpression; O-Glycosylation
AbstractThe accumulation of metabolic by-products remains a critical challenge in the development of mammalian cells culture processes as it impacts cellular growth, productivity and product quality. Although the overexpression of the PYC2 gene was shown to significantly improve the nutrient metabolism efficiency of mammalian cells, its impact on recombinant protein quality has not been investigated yet. In this study, we assess the effect of this metabolic engineering strategy on the quality of a recombinant therapeutic glycoprotein, the human interferon α2b (IFNα2b). As inferred from densitometry analysis of SDS-PAGE gels, PYC2-overexpressing cells sustained a higher percentage of intact glycosylated IFNα2b at the late stage of batch cultures, which was correlated with prolonged viability and reduced accumulation of waste metabolites. Contrarily to the IFNα2b produced by the PYC2 cells, LC–MS analysis confirmed the presence of less glycosylated IFNα2b as well as the occurrence of proteolytic cleavage in the IFNα2b produced in the parental cells. Taken together, these results indicate that PYC2-overexpression in mammalian cells leads to extended favorable conditions for glycosylation and offer an attractive approach to mass-produce high-quality recombinant proteins.
Publication date
PublisherElsevier
LanguageEnglish
AffiliationHuman Health Therapeutics; National Research Council Canada
Peer reviewedYes
NRC numberNRC-HHT-53316
NPARC number23001898
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Record identifiere9bb903b-f264-4ff3-8665-92ee31fa4367
Record created2017-05-09
Record modified2017-05-09
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