Highly sensitive CE-ESI-MS analysis of N-glycans from complex biological samples

Guinevere S.M. Lageveen-Kammeijer; Noortje de Haan; Pablo Mohaupt; Sander Wagt; Mike Filius; Jan Nouta; David Falck; Manfred Wuhrer

doi:10.1038/s41467-019-09910-7

Highly sensitive CE-ESI-MS analysis of N-glycans from complex biological samples

Guinevere S.M. Lageveen-Kammeijer^*, Noortje de Haan, Pablo Mohaupt, Sander Wagt, Mike Filius, Jan Nouta, David Falck, Manfred Wuhrer

^*Corresponding author for this work

BN/Chirlmin Joo Lab

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Abstract

The in-depth, high-sensitivity characterization of the glycome from complex biological samples, such as biofluids and tissues, is of utmost importance in basic biological research and biomarker discovery. Major challenges often arise from the vast structural diversity of glycans in combination with limited sample amounts. Here, we present a method for the highly sensitive characterization of released N-glycans by combining a capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) approach with linkage-specific derivatization of sialic acids and uniform cationic reducing end labelling of all glycans. This method allows the analysis of glycans at the attomole level, provides information on sialic acid isomers and enables the in-depth characterization of complex samples, even when available in minute amounts.

Original language	English
Article number	2137
Number of pages	8
Journal	Nature Communications
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41467-019-09910-7
Publication status	Published - 2019

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s41467-019-09910-7Final published version, 1.71 MBLicence: CC BY

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AU - Lageveen-Kammeijer, Guinevere S.M.

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AU - Mohaupt, Pablo

AU - Wagt, Sander

AU - Filius, Mike

AU - Nouta, Jan

AU - Falck, David

AU - Wuhrer, Manfred

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AB - The in-depth, high-sensitivity characterization of the glycome from complex biological samples, such as biofluids and tissues, is of utmost importance in basic biological research and biomarker discovery. Major challenges often arise from the vast structural diversity of glycans in combination with limited sample amounts. Here, we present a method for the highly sensitive characterization of released N-glycans by combining a capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) approach with linkage-specific derivatization of sialic acids and uniform cationic reducing end labelling of all glycans. This method allows the analysis of glycans at the attomole level, provides information on sialic acid isomers and enables the in-depth characterization of complex samples, even when available in minute amounts.

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Highly sensitive CE-ESI-MS analysis of N-glycans from complex biological samples

Abstract

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