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.
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Acknowledgments
The authors acknowledge funding from EU FP7 under Marie Curie Actions, grant references FP7-PEOPLE-2013-ITN-608381 and FP7-PEOPLE-2012-ITN-316929, and Science Foundation Ireland, grant reference 11/SIRG/B107.
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Mittermayr, S., Albrecht, S., Váradi, C., Millán-Martín, S., Bones, J. (2017). Stable Isotope Quantitative N-Glycan Analysis by Liquid Separation Techniques and Mass Spectrometry. In: Espina, V. (eds) Molecular Profiling. Methods in Molecular Biology, vol 1606. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6990-6_23
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DOI: https://doi.org/10.1007/978-1-4939-6990-6_23
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