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

Wei 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

Wei Wang, G S M Lageveen-Kammeijer^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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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.

Original language	English
Pages (from-to)	143-162
Number of pages	20
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	2531
DOIs	https://doi.org/10.1007/978-1-0716-2493-7_10
Publication status	Published - 2022
Externally published	Yes

Keywords

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

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title = "High-Sensitivity Glycoproteomic Analysis of Biological Samples by CZE-ESI-MS",

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

author = "Wei Wang and Lageveen-Kammeijer, {G S M}",

note = "{\textcopyright} 2022. Springer Science+Business Media, LLC, part of Springer Nature.",

year = "2022",

doi = "10.1007/978-1-0716-2493-7_10",

language = "English",

volume = "2531",

pages = "143--162",

journal = "Methods in molecular biology (Clifton, N.J.)",

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T1 - High-Sensitivity Glycoproteomic Analysis of Biological Samples by CZE-ESI-MS

AU - Wang, Wei

AU - Lageveen-Kammeijer, G S M

PY - 2022

Y1 - 2022

N2 - 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.

AB - 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.

KW - Electrophoresis, Capillary/methods

KW - Proteins

KW - Proteomics/methods

KW - Spectrometry, Mass, Electrospray Ionization/methods

KW - Tandem Mass Spectrometry/methods

KW - Trypsin

U2 - 10.1007/978-1-0716-2493-7_10

DO - 10.1007/978-1-0716-2493-7_10

M3 - Article

C2 - 35941484

SN - 1064-3745

VL - 2531

SP - 143

EP - 162

JO - Methods in molecular biology (Clifton, N.J.)

JF - Methods in molecular biology (Clifton, N.J.)

ER -

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

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Keywords

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