Application of the 50% hydrazine solution method for O-glycans release, their chemical labeling, and HPLC separation

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.