Sialic acid linkage differentiation of glycopeptides using capillary electrophoresis – electrospray ionization – mass spectrometry

Kammeijer, Guinevere S.M.; Jansen, Bas C.; Kohler, Isabelle; Heemskerk, Anthonius A.M.; Mayboroda, Oleg A.; Hensbergen, Paul J.; Schappler, Julie; Wuhrer, Manfred

doi:10.1038/s41598-017-03838-y

Scientific article

English

Sialic acid linkage differentiation of glycopeptides using capillary electrophoresis – electrospray ionization – mass spectrometry

Contributors

Kammeijer, Guinevere S.M.; Jansen, Bas C.; Kohler, Isabelle; Heemskerk, Anthonius A.M.; Mayboroda, Oleg A.; Hensbergen, Paul J.; Schappler, Julie; Wuhrer, Manfred

Published in

Scientific Reports, vol. 7, no. 3733, p. 1-10

Publication date

2017

Abstract

Sialylation is a glycosylation feature that occurs in different linkages at the non-reducing end of a glycan moiety, the linkage isomers are often differentially associated with various biological processes. Due to very similar physico-chemical properties, the separation of isomeric sialylated glycopeptides remains challenging but of utmost importance in the biomedicine and biotechnology, including biomarker discovery, glyco-engineering and biopharmaceutical characterization. This study presents the implementation of a high-resolution separation platform based on capillary electrophoresis – mass spectrometry (CE–MS) allowing for the selective analysis of α2,3- and α2,6-sialylated glycopeptides. These differentially linked glycopeptides showed an identical fragmentation pattern (collision induced dissociation) but different electrophoretic mobilities, allowing for baseline separation of the different linkages without the need for an extensive sample preparation. The different migration behavior between the two moieties was found to correlate with differences in pK_a values. Using a novel methodology adapted from the so-called internal standard CE approach, a relative difference of 3.4·10⁻² in pK_a unit was determined. This approach was applied for the analysis of tryptic glycopeptides of prostate specific antigen, which shows highly complex and heterogeneous glycosylation. The developed platform therefore appears attractive for the identification of differentially linked sialic acids that may be related to pathological conditions.

Keywords

Carbohydrate Conformation
Electrophoresis, Capillary
Glycopeptides / chemistry
N-Acetylneuraminic Acid / chemistry
Spectrometry, Mass, Electrospray Ionization

UNIGE affiliation entities

Faculté des sciences / Section des sciences pharmaceutiques

UNIGE research groups

Sciences analytiques

Funding

European Commission - Methods for high-throughput (HTP) analysis of protein glycosylation [278535]
European Commission - Diagnostic and prognostic biomarkers for inflammatory bowel disease IBD-BIOM [305479]
The Netherlands Genomic Initiative - Horizon Programme Zenith [93511033]
Swiss National Science Foundation - Longitudinal exploratory metabolomics of migraine in transgenic mouse models [155633]

Citation (ISO format)

KAMMEIJER, Guinevere S.M. et al. Sialic acid linkage differentiation of glycopeptides using capillary electrophoresis – electrospray ionization – mass spectrometry. In: Scientific Reports, 2017, vol. 7, n° 3733, p. 1–10. doi: 10.1038/s41598-017-03838-y

Article (Published version)

CC BY-4.0

Appendix

Identifiers

PID : unige:98697
DOI : 10.1038/s41598-017-03838-y
PMID : 28623326
PMCID : PMC5473812

Journal ISSN

2045-2322

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Creation

24/10/2017 09:45:00

First validation

24/10/2017 09:45:00

Update

23/12/2025 14:41:06

Status update

23/12/2025 14:41:06

Last indexation

23/12/2025 14:41:18

Sialic acid linkage differentiation of glycopeptides using capillary electrophoresis – electrospray ionization – mass spectrometry

Contributors

Published in

Publication date

Abstract

Keywords

UNIGE affiliation entities

UNIGE research groups

Funding

Citation (ISO format)

Main files (1)

Secondary files (1)

Identifiers

Journal ISSN

Technical informations

Creation

First validation

Update

Status update

Last indexation