Glycan labeling strategies and their use in identification and quantification.
Most methods for the analysis of oligosaccharides from biological sources require a glycan derivatization step: glycans may be derivatized to introduce a chromophore or fluorophore, facilitating detection after chromatographic or electrophoretic separation. Derivatization can also be applied to link charged or hydrophobic groups at the reducing end to enhance glycan separation and mass-spectrometric detection. Moreover, derivatization steps such as permethylation aim at stabilizing sialic acid residues, enhancing mass-spectrometric sensitivity, and supporting detailed structural characterization by (tandem) mass spectrometry. Finally, many glycan labels serve as a linker for oligosaccharide attachment to surfaces or carrier proteins, thereby allowing interaction studies with carbohydrate-binding proteins. In this review, various aspects of glycan labeling, separation, and detection strategies are discussed.
Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, P.O. Box 9600, 2300RC Leiden, The Netherlands.
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Ruhaak
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Zauner
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Huhn
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Bruggink
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Deelder
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Wuhrer
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry; D002845:Chromatography; D004586:Electrophoresis; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry; D011134:Polysaccharides / Q000737:chemistry* / Q000302:isolation & purification*; D011485:Protein Binding; D013194:Staining and Labeling / Q000379:methods*
D002352:Carrier Proteins; D005456:Fluorescent Dyes; D011134:Polysaccharides