Stable Isotope Quantitative N-Glycan Analysis by Liquid Separation Techniques and Mass Spectrometry

Methods in Molecular Biology
Stefan MittermayrJonathan Bones

Abstract

Liquid phase separation analysis and subsequent quantitation remains a challenging task for protein-derived oligosaccharides due to their inherent structural complexity and diversity. Incomplete resolution or co-detection of multiple glycan species complicates peak area-based quantitation and associated statistical analysis when optical detection methods are used. The approach outlined herein describes the utilization of stable isotope variants of commonly used fluorescent tags that allow for mass-based glycan identification and relative quantitation following separation by liquid chromatography (LC) or capillary electrophoresis (CE). Comparability assessment of glycoprotein-derived oligosaccharides is performed by derivatization with commercially available isotope variants of 2-aminobenzoic acid or aniline and analysis by LC- and CE-mass spectrometry. Quantitative information is attained from the extracted ion chromatogram/electropherogram ratios generated from the light and heavy isotope clusters.

Citations

Aug 14, 2019·Mass Spectrometry Reviews·Fabio P Gomes, John R Yates
Oct 20, 2019·Nanomaterials·Csaba VáradiBéla Viskolcz

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