Enrichment strategies for glycopeptides.
In order to understand glycoprotein functionality, information on the structure of both the core proteins and the glycan moieties is necessary. From a practical viewpoint, glycopeptides rather than whole glycoproteins are the general targets for structural analysis, which is primarily carried out by employing mass spectrometry (MS). Using the "glycoproteomics" concept, several techniques have recently been developed to allow the preparation of a series of reference glycopeptides. In this chapter, we describe two selective capturing methods for glycopeptides, i.e., lectin-affinity chromatography and polysaccharide hydrophilic affinity physicochemical chromatography. The combined use of these methods effectively removes non-glycosylated peptides, the inclusion of which substantially interferes with glycopeptide ionization in MS analysis.
Ito
Shigeyasu
S
Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology, Ibaraki, Japan.
Hayama
Ko
K
Hirabayashi
Jun
J
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D004798:Enzymes / Q000378:metabolism; D054794:Glycomics / Q000379:methods*; D006020:Glycopeptides / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D008954:Models, Biological; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D004798:Enzymes; D006020:Glycopeptides