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Enzymatic Sequence Analysis of N-Glycans by Exoglycosidase Cleavage and Mass Spectrometry – detection of Lewis X Structures

  • Christoph Kannicht
  • Detlef Grunow
  • Lothar Lucka
Part of the Methods in Molecular Biology™ book series (MIMB, volume 446)

Summary

Enzymatic sequencing of oligosaccharides gives structural information on sequence of monosaccharides and type of linkage within the oligosaccharide chain. This data can be obtained by stepwise enzymatic digestion of a single, isolated oligosaccharide using individual or mixtures of specific exoglycosidases. N-glycans have to be fractionated from mixtures prior to sequence analysis to assign this type of structural information to a specific glycan. Enzymatic sequencing can be applied to oligosaccharide mixtures as well to evaluate the occurrence of distinct oligosac-charide motives of functional and/or structural interest.

Here we describe the application of enzymatic sequence analysis to a mixture of N-glycans released from α1-acid glycoprotein. The experimental conditions are optimized for detection of possible Lewis X structures after stepwise exoglycosi-dase digestion by MALDI-TOF mass spectrometry. However, the described method is generally applicable to analyze other structural properties of N-glycans by use of the respective specific exoglycosidases.

Key Words

Enzymatic sequencing N-glycan exoglycosidase MALDI-TOF Lewis x oligosaccharide structure 

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Copyright information

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

Authors and Affiliations

  • Christoph Kannicht
    • 1
  • Detlef Grunow
    • 2
  • Lothar Lucka
    • 2
  1. 1.Molecular Biochemistry DepartmentOctapharma Research and DevelopmentBerlinGermany
  2. 2.Charite Universitätsmedizin BerlinInstitute for Biochemistry and Molecular BiologyBerlinGermany

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