Abstract
The combination of porous graphitized carbon (PGC) liquid chromatography (LC) with mass spectrometric (MS) detection probably constitutes the most elaborate single stage analysis for isomer-specific N-glycan analysis. Here, we describe sample preparation and analysis procedures for the identification of released N-glycans using PGC-LC-ESI-MS and MS/MS.
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References
Pabst M, Altmann F (2011) Glycan analysis by modern instrumental methods. Proteomics 11:631–643
Pabst M, Bondili JS, Stadlmann J et al (2007) Mass + retention time = structure: a strategy for the analysis of N-glycans by carbon LC-ESI-MS and its application to fibrin N-glycans. Anal Chem 79:5051–5057
Stadlmann J, Pabst M, Kolarich D et al (2008) Analysis of immunoglobulin glycosylation by LC-ESI-MS of glycopeptides and oligosaccharides. Proteomics 8:2858–2871
Pabst M, Grass J, Toegel S et al (2012) Isomeric analysis of oligomannosidic N-glycans and their dolichol-linked precursors. Glycobiology 22:389–399
Everest-Dass AV, Jin D, Thaysen-Andersen M et al (2012) Comparative structural analysis of the glycosylation of salivary and buccal cell proteins: innate protection against infection by Candida albicans. Glycobiology 22:1465–1479
Jensen PH, Karlsson NG, Kolarich D et al (2012) Structural analysis of N- and O-glycans released from glycoproteins. Nat Protoc 7:1299–1310
Tretter V, Altmann F, Marz L (1991) Peptide-N4-(N-acetyl-beta-glucosaminyl)-asparagine amidase F cannot release glycans with fucose attached alpha 1→3 to the asparagine-linked N-acetylglucosamine residue. Eur J Biochem 199:647–652
Kolarich D, Altmann F (2000) N-Glycan analysis by matrix-assisted laser desorption/ionization mass spectrometry of electrophoretically separated nonmammalian proteins: application to peanut allergen Ara h 1 and olive pollen allergen Ole e 1. Anal Biochem 285:64–75
Harvey DJ (2005) Structural determination of N-linked glycans by matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry. Proteomics 5:1774–1786
Everest-Dass AV, Abrahams JL, Kolarich D et al (2013) Structural feature ions for distinguishing N- and O-Linked glycan isomers by LC-ESI-IT MS/MS. J Am Soc Mass Spectrom 24:895–906
Everest-Dass AV, Kolarich D, Campbell MP et al (2013) Tandem mass spectra of glycan substructures enable the multistage mass spectrometric identification of determinants on oligosaccharides. Rapid Commun Mass Spectrom 27:931–939
Kolarich D, Rapp E, Struwe WB et al (2013) The minimum information required for a glycomics experiment (MIRAGE) project: improving the standards for reporting mass-spectrometry-based glycoanalytic data. Mol Cell Proteomics 12:991–995
York WS, Agravat S, Aoki-Kinoshita KF et al (2014) MIRAGE: the minimum information required for a glycomics experiment. Glycobiology 24:402–406
Plummer TH Jr, Phelan AW, Tarentino AL (1987) Detection and quantification of peptide-N4-(N-acetylbeta-glucosaminyl)asparagine amidase. Eur J Biochem 163:167–173
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Kolarich, D., Windwarder, M., Alagesan, K., Altmann, F. (2015). Isomer-Specific Analysis of Released N-Glycans by LC-ESI MS/MS with Porous Graphitized Carbon. In: Castilho, A. (eds) Glyco-Engineering. Methods in Molecular Biology, vol 1321. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2760-9_29
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DOI: https://doi.org/10.1007/978-1-4939-2760-9_29
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2759-3
Online ISBN: 978-1-4939-2760-9
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