Strategy for the investigation of O-linked oligosaccharides from mucins based on the separation into neutral, sialic acid- and sulfate-containing species |
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Authors: | Niclas G Karlsson Hasse Karlsson and Gunnar C Hansson |
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Institution: | (1) Department of Medical Biochemistry, University of Göteborg, Medicinaregatan 9, 413 90 Gothenburg, Sweden |
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Abstract: | A method for the separation of O-linked oligosaccharides into neutral, sialic acid-containing and sulfated species was applied to oligosaccharides released by alkaline borohydride from mucin glycopeptides from porcine small intestine. The released mixture of reduced oligosaccharides was applied to an anion exchange column, and the neutral oligosaccharides were collected as the unretarded fraction. A mixture of dimethyl sulfoxide and iodomethane was passed through the column to convert the sialic acid-containing oligosaccharides into methyl esters that were eluted and converted to methyl amides by methyl amine. Finally the sulfated oligosaccharide fraction was eluted with salt. The neutral and the derivatized sialic acid-containing oligosaccharides were analysed by gas chromatography-mass spectrometry after permethylation and the sulfated oligosaccharide fraction was analysed by high performance anion exchange chromatography.Abbreviations GC
gas chromatography
- GC/MS
gas chromatography-mass spectrometry
- HPAEC-PAD
high performance anion exchange chromatography-pulsed amperometric detection
- Hex
hexose
- HexNAc
N-acetyl hexosamine
- HexNAcol
N-acetyl hexosaminitol
- Fuc
Fucose
- NeuAc
N-acetyl neuraminic acid
- NeuGc
N-glycolyl neuraminic acid |
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Keywords: | Gas chromatography-mass spectrometry oligosaccharide mucin high performance anion exchange chromatography sialic acid sulfate |
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