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Methods Mol Biol. 2013;951:145-69. doi: 10.1007/978-1-62703-146-2_11.

Separation and identification of glycoforms by capillary electrophoresis with electrospray ionization mass spectrometric detection.

Author information

1
Department of Chemical and Biological Sciences, "Aurel Vlaicu" University of Arad, Timisoara, Romania. alina.zamfir@uav.ro

Abstract

Capillary electrophoresis (CE) is a resourceful and versatile separation method for the analysis of complex carbohydrate mixtures. In combination with electrospray ionization (ESI) mass spectrometry (MS), CE enables fast, sensitive, and efficient separations for the accurate identification of a large variety of glycoform mixture types. In this chapter several reliable off- and on-line CE-based methods for the analysis of glycoforms with ESI MS/MS are presented. The first part of this chapter is dedicated to the application of off-line CE/ESI MS to complex mixtures of O-glycopeptides and mixtures of proteoglycan-derived O-glycans, i.e., glycosaminoglycans such as depolymerized hybrid chains of chondroitin sulfate (CS) and dermatan sulfate (DS). Procedures for off-line fractionation of these heterogeneous mixtures followed by ESI MS screening and sequencing of single glycoforms by collision-induced dissociation (CID) at low energies are also described. Ample sections are further devoted to on-line CE/ESI MS technique and its application to separation and identification of O-glycopeptides and CS/DS oligosaccharides. The concept and construction principles of two different sheathless CE/ESI MS interfaces together with the protocols to be applied for successful on-line analysis of O-glycopeptides and CS/DS oligosaccharides are presented in details in the last part of the chapter.

PMID:
23296530
DOI:
10.1007/978-1-62703-146-2_11
[Indexed for MEDLINE]

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