Liquid Chromatography and Capillary Electrophoresis Analysis of 2AA-Labeled O-Glycans.
Methods Mol Biol
; 2763: 171-185, 2024.
Article
em En
| MEDLINE
| ID: mdl-38347410
ABSTRACT
To reveal O-glycan structures in mucins, it is necessary to release covalently bound O-glycans from the polypeptide backbone and derivatize to a form suitable for structural analysis. Various derivatization methods can now be applied in the analysis of O-glycans following the development of O-glycan release methods. Among the many derivatization methods available, we prefer to use fluorescent labeling with 2-aminobenzoic acid (anthranilic acid, 2AA). 2AA-labeled O-glycans can be detected with high sensitivity using liquid chromatography fluorescence detection (LC-FD) analysis because of the strong fluorescence. In addition, as 2AA has a carboxyl group that carries a negative charge, 2AA-labeled O-glycans can be analyzed with high sensitivity in negative ion mode mass spectrometry. Furthermore, because the negative charge of 2AA provides a driving force for electrophoresis, 2AA-labeled O-glycans can be analyzed using capillary electrophoresis (CE) and capillary affinity electrophoresis. High detection sensitivity and versatility are key advantages of the 2AA-labeling method. Here we present our preferred LC-FD and CE analytical methods for 2AA-labeled O-glycans.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Polissacarídeos
/
Eletroforese Capilar
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Japão