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Updated: Jan 22, 2026

Semi-Quantitative Analysis of Peptidoglycan by Liquid Chromatography Mass Spectrometry and Bioinformatics
Published on: October 13, 2020
Enzymatic Sequence Analysis of N-Glycans by Exoglycosidase Cleavage and Mass Spectrometry: Detection of Lewis X
Christoph Kannicht1, Detlef Grunow2, Lothar Lucka3
1Recombinant Research and Development, Octapharma Biopharmaceuticals GmbH, Heidelberg, Germany. christoph@kannicht.net.
Abstract:
Enzymatic sequencing of oligosaccharides provides 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 as well be applied to oligosaccharide mixtures to evaluate the occurrence of distinct oligosaccharide 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 exoglycosidase digestion by MALDI-TOF mass spectrometry. However, the described method is generally applicable to analyze other structural properties of N-glycans using (respective) specific exoglycosidases.
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