Highly sensitive CE-ESI-MS analysis of N-glycans from complex biological samples
Guinevere S.M. Lageveen-Kammeijer (Leiden University Medical Center)
Noortje de Haan (Leiden University Medical Center)
Pablo Mohaupt (Leiden University Medical Center)
Sander Wagt (Leiden University Medical Center)
Mike Filius (TU Delft - BN/Chirlmin Joo Lab, Leiden University Medical Center)
Jan Nouta (Leiden University Medical Center)
David Falck (Leiden University Medical Center)
Manfred Wuhrer (Leiden University Medical Center)
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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.
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