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Isomer-Specific Analysis of Released N-Glycans by LC-ESI MS/MS with Porous Graphitized Carbon

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Glyco-Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1321))

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

  1. Pabst M, Altmann F (2011) Glycan analysis by modern instrumental methods. Proteomics 11:631–643

    Article  CAS  PubMed  Google Scholar 

  2. 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

    Article  CAS  PubMed  Google Scholar 

  3. 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

    Article  CAS  PubMed  Google Scholar 

  4. Pabst M, Grass J, Toegel S et al (2012) Isomeric analysis of oligomannosidic N-glycans and their dolichol-linked precursors. Glycobiology 22:389–399

    Article  CAS  PubMed  Google Scholar 

  5. 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

    Article  CAS  PubMed  Google Scholar 

  6. Jensen PH, Karlsson NG, Kolarich D et al (2012) Structural analysis of N- and O-glycans released from glycoproteins. Nat Protoc 7:1299–1310

    Article  CAS  PubMed  Google Scholar 

  7. 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

    Article  CAS  PubMed  Google Scholar 

  8. 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

    Article  CAS  PubMed  Google Scholar 

  9. Harvey DJ (2005) Structural determination of N-linked glycans by matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry. Proteomics 5:1774–1786

    Article  CAS  PubMed  Google Scholar 

  10. 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

    Article  CAS  PubMed  Google Scholar 

  11. 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

    Article  CAS  PubMed  Google Scholar 

  12. 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

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. York WS, Agravat S, Aoki-Kinoshita KF et al (2014) MIRAGE: the minimum information required for a glycomics experiment. Glycobiology 24:402–406

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. 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

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Friedrich Altmann .

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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

  • eBook Packages: Springer Protocols

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