Abstract
We developed a new method for MALDI-TOF MS detection of N-glycans derived from human serum. The synergistic combination of microwave-assisted Girard T derivatization, solid-phase extraction desalting, and an ionic liquid matrix (2, 5-dihydroxybenzoic acid/aniline) (GT-SPE-DHB/An) allowed of more sensitive N-glycans detection than a conventional ionic liquid matrix in MALDI-TOF MS. The superior sensitivity of our method was confirmed by the number of assigned N-glycans in 900–2,000 m/z range. Using our GT-SPE-DHB/An method, we were successfully able to assign 31 glycans. However, with the established method, i.e., DHB/An method, only 15 glycans were assigned. To the best of our knowledge, this GT-SPE-DHB/An method is the first to combine cationic derivatization of N-glycan and ionic liquid matrix for N-glycan analysis in MALDI-TOF MS.
Highlights
N-Linked glycans are involved in many biological processes such as protein folding, cell-cell interaction, and immune response [1, 2]
N-glycosylation changes in the serum glycome are related to cancer and inflammation [3], and serum glycome profiling can serve as tumor biomarkers [4]. erefore, N-glycan analysis in human serum is of central importance in biomarker discovery [5] and cancer diagnostics [6]
Glycan analysis based on matrix-assisted laser desorption/ionization- (MALDI-) time of flight (TOF) mass spectrometry (MS) has several advantages: low sample consumption, high sensitivity, and simple sample preparation
Summary
N-Linked glycans are involved in many biological processes such as protein folding, cell-cell interaction, and immune response [1, 2]. Erefore, N-glycan analysis in human serum is of central importance in biomarker discovery [5] and cancer diagnostics [6]. Despite their importance, the following characteristics of glycans make glycan analysis more challenging: heterogeneity of N-glycosylation [7] and absence of a unified database for glycome structure [8]. Glycan analysis based on matrix-assisted laser desorption/ionization- (MALDI-) time of flight (TOF) mass spectrometry (MS) has several advantages: low sample consumption, high sensitivity, and simple sample preparation. Girard’s reagent T ((carboxymethyl)trimethylammonium chloride hydrazide, GT) and P (1-(carboxymethyl)pyridinium chloride hydrazide, GP) were used to improve the ionization efficiency of glycans in MALDI and ESI mass spectrometry [15]. ILMs have gained increasing attention because of their advantages in vacuum stability and signal-to-noise
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