Application of the 50% Hydrazine Solution Method for O-Glycans Release, their Chemical Labeling, and HPLC Separation

D. G. Kisiel Department of Medical Chemistry, Medical University of Białystok, Mickiewicza, Białystok, Poland

I. Radziejewska Department of Medical Chemistry, Medical University of Białystok, Mickiewicza, Białystok, Poland

A. Gindzieński Department of Medical Chemistry, Medical University of Białystok, Mickiewicza, Białystok, Poland

ABSTRACT

Mucins are high molecular mass glycoproteins with oligosaccharides O-bonded to the protein core. β-elimination is the most popular method used for releasing of O-glycans. However to such released glycoforms it is difficult to introduce a label to amplify a signal for oligosaccharide detection.

In our study we used a combination of the β-elimination and hydrazinolysis methods. Released glycoforms were labeled with para-amino benzooic acid ethyl ester (ABBE) and fractionated on HPLC column.

This combined procedure seems to be a good tool for O-glycans analysis.

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Application of the 50% Hydrazine Solution Method for O-Glycans Release, their Chemical Labeling, and HPLC Separation

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Application of the 50% Hydrazine Solution Method for O-Glycans Release, their Chemical Labeling, and HPLC Separation

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