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Exploratory Approach of Serum N-Glycan Profiling by Mass Spectrometry

Authors: Rahman, Salina Abdul; Siti Aishah Abdul Wahab; Ngu Lock Hock;

Exploratory Approach of Serum N-Glycan Profiling by Mass Spectrometry

Abstract

Introduction Congenital disorders of glycosylation (CDG) are among the inherited metabolic diseases caused by a defect in protein glycosylation. We have used our modified FASP protocol (‘glycoFASP’) to determine the structures of normal and CDG Type 1 patient Nglycans. Objective Develop a simple FANGS3 protocol to isolate and analyze glycoprotein glycans from human serum. • Apply the glycoFASP method to compare N-glycan profiles between normal control and CDG Type 1 patient. Methodology [Refer to Poster]. Results glycoFASP-released glycans from normal control and CDG Type 1 patient were analysed using MALDI –TOF/TOF MS are shown in Figure 1. The spectrum shows glycans signal (m/z 1419, 1581 and 1744 ) are not observed from CDG Type 1 patient when compared to normal control spectrum. Normal control also produce non fucosylated (m/z 1501) and fucosylated (m/z 1485,1648 and 1810) complex biantennary, bisecting GlcNAc (m/z 1689, 1851, 2013 and 2175) and sialylated (m/z 1955, 2101 and 2246) glycans. Interestingly, CDG Type 1 spectrum also show non fucosylated (m/z 1501) and fucosylated (m/z 1485,1648 and 1810) complex bi-antennary, sialylated (m/z 1955,2101 and 2246) and bisecting GlcNAc (m/z 1689, 1851 and 2013) glycans lacking (m/z 2175 ). These N-glycans signal are markedly lower compared to normal control. Conclusion The difference between normal control and CDG Type 1 glycan structures has been demonstrated. • The protocol is less laborious and reduces time in sample preparation before MALDI analysis. • The protocol works efficiently and convenience for low volume of sample.

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Keywords

Serum N-Glycan Profiling, Mass Spectrometry

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This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
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