Molecules · 2014 · 37 citations · 20 references
Life SciencesBiochemistryMolecular BiologyChromatographic AnalysisGlycobiologyProteomicsAnalytical ChromatographyChromatographyGlycan AnalysisGlycan-protein InteractionsLiquid ChromatographyProtein GlycosylationAnalytical Method DevelopmentColumnsConjugationGlycan Derivatization For Structural AnalysisGlycopeptide AnalysisGlycoprotein AnalysisIntact Glycoprotein AnalysisProtein Glycosylation Analysis
In this study, an HPLC HILIC-UV method was developed for the analysis of intact neo-glycoproteins. During method development the experimental conditions evaluated involved different HILIC columns (TSKgel Amide-80 and ZIC-pHILIC), and water-acetonitrile mixtures containing various types of acids and salts. The final selected method was based on a TSKgel Amide-80 column and a mobile phase composed of acetonitrile and water both containing 10 mM HClO4. The influence of temperature and sample preparation on the chromatographic performances of the HILIC method was also investigated. The method was applied to the separation of neo-glycoproteins prepared starting from the model protein RNase A by chemical conjugation of different glycans. Using the method here reported it was possible to monitor by UV detection the glycosylation reaction and assess the distribution of neo-glycoprotein isoforms without laborious sample workup prior to analysis.
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Jonathan Bones, Stefan Mittermayr, Niaobh O’Donoghue et al. · Analytical Chemistry · 2010 · 167 citations
Synthetically defined glycoprotein vaccines: current status and future directions
Roberto Adamo, Alberto Nilo, Bastien Castagner et al. · Chemical Science · 2013 · 156 citations · Full text