High-Sensitivity Glycoproteomic Analysis of Biological Samples by CZE-ESI-MS

W Wang; G S M Lageveen-Kammeijer

doi:10.1007/978-1-0716-2493-7_10

High-Sensitivity Glycoproteomic Analysis of Biological Samples by CZE-ESI-MS

Methods Mol Biol. 2022:2531:143-162. doi: 10.1007/978-1-0716-2493-7_10.

Authors

W Wang¹, G S M Lageveen-Kammeijer²

Affiliations

¹ Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
² Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands. g.s.m.kammeijer@lumc.nl.

PMID: 35941484
DOI: 10.1007/978-1-0716-2493-7_10

Abstract

Capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) is a powerful tool for the characterization and identification of the macro- and microheterogeneity of a glycoprotein in a bottom-up approach. This chapter describes in detail the sample preparation procedures using a purified biological sample, prostate-specific antigen, as a model protein, including proteolytic digestion (trypsin). In addition, insights are provided into the strengths of using capillary electrophoresis for obtaining isomer separation of differently linked sialic acids. Lastly, approaches and potential pitfalls for the integration and quantitation of glycopeptide signals from the obtained CZE-MS data are discussed.

Keywords: Bottom-up proteomics; Capillary electrophoresis-mass spectrometry (CE-MS); Data analysis; Glycoform profiling; Glycopeptide analysis; Glycoproteomics; Prostate-specific antigen (PSA).

MeSH terms

Electrophoresis, Capillary / methods
Proteins
Proteomics* / methods
Spectrometry, Mass, Electrospray Ionization / methods
Tandem Mass Spectrometry* / methods
Trypsin

Substances

Proteins
Trypsin