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Separation of N-Glycans by HPLC

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Post-translational Modifications of Proteins

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 446))

Summary

Most glycoproteins carry a very heterogeneous mixture of oligosaccharides and even a single glycosylation site of a pure glycoprotein is often heterogeneously glycosylated. The structural diversity of oligosaccharides arises from linkage variants, from differences in the size and number of charges of glycans, and from differences in the monosaccharide composition of glycans. Fortunately, the biosynthetic pathway is subject to certain restrictions, so that structural diversity is limited and amenable to laboratory investigation. Different approaches have been developed to the structural characterization of oligosaccharides, including nuclear magnetic resonance (NMR), mass-spectrometry, linkage analysis by gas chromatography-mass spectrometry (GC-MS), sequence analysis using specific exoglycosidases and others, but a crucial part of these strategies is the separation of the glycan mixture into homogeneous glycan fractions. In this chapter some high-performance liquid chromatography (HPLC)-techniques are described for the isolation of oligosaccharides, in particular N-linked glycans.

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Author information

Authors and Affiliations

  1. Dynavax Technologies, Berkeley, CA, USA

    Martin Gohlke

  2. Zentralinstitut fur Laboratoriumsmedizin und Pathobiochemie, Charité-Universitatsmedizin Berlin, Berlin, Germany

    Véronique Blanchard

Authors
  1. Martin Gohlke
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  2. Véronique Blanchard
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Editor information

Editors and Affiliations

  1. Molecular Biochemistry Department, Octapharma Research and Development, Berlin, Germany

    Christoph Kannicht

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Gohlke, M., Blanchard, V. (2008). Separation of N-Glycans by HPLC. In: Kannicht, C. (eds) Post-translational Modifications of Proteins. Methods in Molecular Biology™, vol 446. Humana Press. https://doi.org/10.1007/978-1-60327-084-7_17

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  • DOI: https://doi.org/10.1007/978-1-60327-084-7_17

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-719-8

  • Online ISBN: 978-1-60327-084-7

  • eBook Packages: Springer Protocols

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