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Fractionation and characterization of sialyl linkage isomers of serum N-glycans by CE-MS.
- Source :
-
Journal of separation science [J Sep Sci] 2022 Sep; Vol. 45 (17), pp. 3348-3361. - Publication Year :
- 2022
-
Abstract
- Structural isomers of sialylated N-glycans contribute to the diversity of the N-glycome and to a range of biological functions. Sialyl linkage isomers can be readily distinguished by mass spectrometry with mass differences between α2,3- and α2,6-linkages generated by a two-step sialic acid linkage-specific alkylamidation. To improve the identification of N-glycans from complex mixtures, we added a delactonization step after the first alkylamidation step, which regenerates negatively charged carboxylic acids on α2,3-sialic acids. N-glycan isomers with α2,3-sialic acids are then fractionated by ion-exchange chromatography prior to the second alkylamidation step. With this modified alkylamidation method, sialylated N-glycans were enriched and stabilized for structural characterization by capillary electrophoresis-mass spectrometry and tandem mass spectrometry. We identified 52 sialylated N-glycan structures, including 107 linkage isomers, in human serum and confirmed the presence of positional isomers of specific sialyl linkage isomers. Due to the reduced sample complexity after ion-exchange fractionation and CE separation, substructural features of N-glycans were rapidly evaluated and included core- and antenna-fucosylation and poly-lactosamine.<br /> (© 2022 Wiley-VCH GmbH.)
Details
- Language :
- English
- ISSN :
- 1615-9314
- Volume :
- 45
- Issue :
- 17
- Database :
- MEDLINE
- Journal :
- Journal of separation science
- Publication Type :
- Academic Journal
- Accession number :
- 35819141
- Full Text :
- https://doi.org/10.1002/jssc.202200223