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• W3087481505 abstract "Characterization of the structural diversity of glycans by liquid chromatography–tandem mass spectrometry (LC–MS/MS) remains an analytical challenge in large-scale glycomics applications because of the presence of heterogeneous composition, ubiquitous isomers, lability of post-translational glycan modifications, and complexity of data interpretation. High-resolution separation of glycan isomers differentiating from positional, linkage, branching, and anomeric structures is often a prerequisite to ensure the comprehensive glycan identification. Here, we developed a straightforward method using self-packed capillary porous graphitic carbon (PGC) columns for nanoflow LC–MS/MS analyses of native glycans released from glycoproteins. The technique enables highly resolved chromatographic separation of over 20 high-mannose glycan isomers in ribonuclease B and a diverse range of hybrid and complex-type sialoglycoforms of fetuin. The distinct structures of anomeric glycans and linkage sialoglycan isomers, α2,3 and α2,6, were identified by the characteristic MS/MS fragment ions. A glycan sequencing strategy utilizing diagnostic ions and complementary fragments specific to branching residues was established to simplify the MS/MS data interpretation of closely related isomeric structures. To promote the PGC-LC–MS/MS-based method for glycome-wide applications, we extended analyses to native sulfoglycans from the egg-propagated and cell culture-derived influenza vaccines and demonstrate the high-resolution separation and structural characterization of underivatized neutral and anionic glycoforms including oligomannosidic glycan anomers, sialoglycan linkage isomers, and regioisomers of afucosylated and fucosylated sulfoglycans containing sulfated-6-GlcNAc and sulfated-4-GalNAc residues." @default.
• W3087481505 created "2020-09-25" @default.
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• W3087481505 date "2020-09-22" @default.
• W3087481505 modified "2023-10-18" @default.
• W3087481505 title "Resolving Isomeric Structures of Native Glycans by Nanoflow Porous Graphitized Carbon Chromatography–Mass Spectrometry" @default.
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• W3087481505 doi "https://doi.org/10.1021/acs.analchem.0c02951" @default.
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• W3087481505 hasPublicationYear "2020" @default.
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