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    Home > Biochemistry News > Biotechnology News > Glycoprotein Analysis by Capillary Zone Electrophoresis-Electrospray Mass Spectrometry

    Glycoprotein Analysis by Capillary Zone Electrophoresis-Electrospray Mass Spectrometry

    • Last Update: 2021-02-26
    • Source: Internet
    • Author: User
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    The study of glycoproteins as intact molecules or as smaller fragments generated by chemical or enzymatic digestion is wellsuited to the technique of capillary zone electrophoresis-electrospray mass spectrometry (CZE-ESMS). Separation of glycoformpopulations of intact glycoproteins or protein digests is possible using buffers compatible with mass spectrometric detection(
    1

    8
    ). CZE-ESMS analysis of intact proteins can provide semiquantitative information about the degree of heterogeneity of theglycoprotein (
    1
    ,
    6
    ,
    7
    ). Enzymatic or chemical digestion of the glycoprotein, followed by CZE-ESMS, gives a direct measure of individual sites ofheterogeneity (
    1

    5
    ). Combined CZE-collision induced dissociation (CID) mass spectrometric experiments (CZE-MS-MS) of digests enables the characterizationof oligosaccharide structures (
    1

    5
    ). In particular, CID of glycopeptides is characterized by fragment ions corresponding to cleavage at each glycosidic bond. The occurrence of specific carbohydrate residues such as hexose (Man, Glc, Gal),
    N
    -acetylhexosamine (GlcNAc, GalNAc), or N-acetylneuraminic acid (NeuNAc) can be monitored by the observation of characteristicoxonium ions at
    m/z
    163, 204, and 292, respectively. More recently, CZE-front-end CID-MS-MS has been shown to be a powerful tool for peptidesequencing of glycopeptides using subpicomole quantities of injected glycoprotein (
    3

    5
    ).
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