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Anal Bioanal Chem. 2017 Feb;409(5):1257-1269. doi: 10.1007/s00216-016-0052-5. Epub 2016 Nov 10.

Efficient recovery of glycosaminoglycan oligosaccharides from polyacrylamide gel electrophoresis combined with mass spectrometry analysis.

Author information

1
CNRS UMR 8587, Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement, 91025, Evry, France.
2
Université Evry-Val-d'Essonne, Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement, F-91025, Evry, France.
3
INSERM U1088, Laboratoire de Biochimie Spécialisée, Centre de Biologie Humaine, CHU Sud, 80054, Amiens Cedex, France.
4
CNRS UMR 8587, Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement, 91025, Evry, France. regis.daniel@univ-evry.fr.
5
Université Evry-Val-d'Essonne, Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement, F-91025, Evry, France. regis.daniel@univ-evry.fr.

Abstract

To promote efficient separation and structural analysis of glycosaminoglycan oligosaccharides, we developed a straightforward method that combined gel electrophoresis and mass spectrometry (MS). Potential limitations of this approach (e.g., low extraction yields and weak compatibility with MS) were resolved by developing an active extraction procedure that yielded a quantitative amount of sulfated oligosaccharides from excised gel bands. The compatibility of obtained oligosaccharides for subsequent MS analysis was ensured using a single, simple clean-up step on a mixed C18/graphite carbon solid-phase column that was fully effective for polymerization degrees ranging from di- to dodecasaccharides. The reported combination of carbohydrates-polyacrylamide gel electrophoresis with MS was successfully applied to glucosamino- (heparin) and galactosamino- (dermantan sulfate) glycans, demonstrating the potential of our method for structural analysis of bioactive sulfated carbohydrates extracted from biological matrices. Graphical Abstract ᅟ.

KEYWORDS:

C-PAGE; Gel extraction; Glycosaminoglycan; Mass spectrometry

PMID:
27832303
DOI:
10.1007/s00216-016-0052-5
[Indexed for MEDLINE]

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