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Liquid chromatography-electrospray mass spectrometry as a tool for the analysis of sulfated oligosaccharides from mucin glycoproteins.

Journal article
Authors Kristina A Thomsson
Niclas G. Karlsson
Gunnar C. Hansson
Published in Journal of chromatography. A
Volume 854
Issue 1-2
Pages 131-9
ISSN 0021-9673
Publication year 1999
Published at Institute of Medical Biochemistry
Pages 131-9
Language en
Links www.ncbi.nlm.nih.gov/entrez/query.f...
Keywords Animals, Carbohydrate Sequence, Chromatography, High Pressure Liquid, methods, Glycoproteins, chemistry, Mass Spectrometry, methods, Molecular Sequence Data, Mucins, chemistry, Oligosaccharides, analysis, Sulfuric Acids, chemistry, Swine
Subject categories Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

Abstract

An approach for analyzing sulfated oligosaccharide alditol mixtures by liquid chromatography-electrospray mass spectrometry (LC-ESI-MS) is described. Two columns, an amino-bonded column and a porous graphitized carbon column (PGC) were used. Oligosaccharides were eluted with linear gradients of acetonitrile and water, with 5 mM ammonium hydrogencarbonate or formate buffers at a basic pH. The methods were evaluated on a mixture of sulfated oligosaccharide alditols prepared from mucin glycoproteins from pig stomach. Results from LC-ESI-MS of the mixture were compared with the structural information obtained by high energy collision fragmentation using fast atom bombardment tandem mass spectrometry (FAB-MS-MS). The separation ability of the two columns was also tested using a more complex mixture of sulfated oligosaccharides from pig colon, where several isomers were detected. The potential use of in-source collision-induced dissociation (CID) to gain sequence information of sulfated oligosaccharides was also evaluated. The major fragment ions obtained by in-source CID of the trisaccharide Hex-3HexNAcol6-HexNAc6-SO3 were sufficient for assigning the oligosaccharide sequence and the position of the sulfate group within the monosaccharide moiety. The LC-ESI-MS approach should be a valuable tool for characterization of mucin glycosylation and alterations during pathological conditions.

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