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129th General Meeting of Korean Chemical Society & Exposition Absolute Quantification of Glycans using Unlabeled Standard and Isotope-labeled Analytes by Mass Spectrometry

Submission Date :
2 / 28 / 2022 , 15 : 53 : 50
Abstract Number :
Presenting Type:
Poster Presentation
Presenting Area :
Analytical Chemistry
Authors :
Jihee Yun, Jae-Min Lim*
Department of Chemistry, Changwon National University, Korea
Assigned Code :
ANAL.P-338 Assigend Code Guideline
Presenting Time :
April 14 (THU) 11:00~13:00
Protein glycosylation is one of the most common post-translational modifications (PTMs). Glycans not only directly participate in cell development and cell differentiation, but also provide genetic or pathological information. It is important to understand changes in glycan expression levels to identify correlations with glycan function and disease. Therefore, many relative quantification techniques have been developed through isotope labeling strategies based on mass spectrometry. However, absolute quantification techniques suffer from many difficulties due to the synthesis of isotopically labeled standards. In this study, Quantitative Analysis using Unlabeled Standard and Isotope-labeled Analytes (QAUSIA) is presented to realize absolute quantification of glycan. We successfully labeled glycans in yeast using 1-13C1 glucose based on Metabolic Isotope Labeling of Polysaccharides with Isotopic Glucose (MILPIG) strategy for isotopic labeling of analytes. Then, it was combined with an unlabeled internal standard to obtain a calibration curve showing excellent linearity and reproducibility based on mass spectrometry. In addition, the applicability in biological conditions was confirmed using tunicamycin, a glycosylation inhibitor, through two internal standards. In conclusion, we indicate that QAUSIA is relevant for establishing a simple, fast, and effective absolute quantitative assay for analyzing absolute glycan information in disease.