A method of purification of partially methylated alditol acetates in the methylation analysis of glycoproteins and glycopeptides

Mark E. Lowe; Bo Nilsson

doi:10.1016/0003-2697(84)90323-3

A method of purification of partially methylated alditol acetates in the methylation analysis of glycoproteins and glycopeptides

Mark E. Lowe, Bo Nilsson

Research output: Contribution to journal › Article

8 Scopus citations

Abstract

A method for methylation analysis of intact glycoproteins is described. Starting with intact glycoprotein, the oligosaccharides are methylated, hydrolyzed, reduced, and acetylated. The partially methylated alditol acetates are then separated from noncarbohydrate contaminants on a silica gel G column. Partially methylated hexitol acetates are eluted from the column with petroleum ether:ethyl acetate (1:1, v v) and partially methylated N-acetylhexosaminitol acetates are subsequently eluted with methanol. Analysis by gas-liquid chromatography/mass spectrometry of the partially methylated alditol acetates shows no interfering contaminants. This method circumvents the need to make pronase glycopeptides and avoids the pitfalls of other methylation procedures.

Original language	English
Pages (from-to)	187-191
Number of pages	5
Journal	Analytical Biochemistry
Volume	136
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(84)90323-3
State	Published - Jan 1984
Externally published	Yes

Keywords

N-linked
glycopeptide
glycoprotein
methylation
oligosaccharide
silica

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10.1016/0003-2697(84)90323-3

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Lowe, M. E., & Nilsson, B. (1984). A method of purification of partially methylated alditol acetates in the methylation analysis of glycoproteins and glycopeptides. Analytical Biochemistry, 136(1), 187-191. https://doi.org/10.1016/0003-2697(84)90323-3

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N2 - A method for methylation analysis of intact glycoproteins is described. Starting with intact glycoprotein, the oligosaccharides are methylated, hydrolyzed, reduced, and acetylated. The partially methylated alditol acetates are then separated from noncarbohydrate contaminants on a silica gel G column. Partially methylated hexitol acetates are eluted from the column with petroleum ether:ethyl acetate (1:1, v v) and partially methylated N-acetylhexosaminitol acetates are subsequently eluted with methanol. Analysis by gas-liquid chromatography/mass spectrometry of the partially methylated alditol acetates shows no interfering contaminants. This method circumvents the need to make pronase glycopeptides and avoids the pitfalls of other methylation procedures.

AB - A method for methylation analysis of intact glycoproteins is described. Starting with intact glycoprotein, the oligosaccharides are methylated, hydrolyzed, reduced, and acetylated. The partially methylated alditol acetates are then separated from noncarbohydrate contaminants on a silica gel G column. Partially methylated hexitol acetates are eluted from the column with petroleum ether:ethyl acetate (1:1, v v) and partially methylated N-acetylhexosaminitol acetates are subsequently eluted with methanol. Analysis by gas-liquid chromatography/mass spectrometry of the partially methylated alditol acetates shows no interfering contaminants. This method circumvents the need to make pronase glycopeptides and avoids the pitfalls of other methylation procedures.

KW - N-linked

KW - glycopeptide

KW - glycoprotein

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KW - oligosaccharide

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