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A rapid sample preparation method for mass spectrometric characterization of N‐linked glycans
Author(s) -
Yu Ying Qing,
Gilar Martin,
Kaska Jennifer,
Gebler John C.
Publication year - 2005
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2067
Subject(s) - chemistry , chromatography , glycan , sample preparation , mass spectrometry , matrix assisted laser desorption/ionization , tandem mass spectrometry , hydrophilic interaction chromatography , derivatization , elution , electrospray , desorption , glycoprotein , high performance liquid chromatography , adsorption , organic chemistry , biochemistry
A rapid method for analysis of glycans of glycoproteins is presented. This method comprised deglycosylation, sample cleanup and matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) analysis of glycans. The enzymatic deglycosylation of N‐linked glycoproteins was enhanced in terms of speed and reproducibility using an enzyme‐friendly surfactant. The released glycans were desalted using a micro‐scale solid phase extraction (SPE) device packed with a hydrophilic interaction chromatography (HILIC) sorbent. Hydrophilic glycans were well retained by SPE, while salts and surfactants were removed from the sample. The glycans were eluted using 25–50 µL of solvent and analyzed directly without derivatization using MALDI‐MS. MALDI quadrupole time‐of‐flight (Q‐Tof) instrumentation was utilized for glycan profiling and structure characterization by tandem mass spectrometry (MS/MS). The presented method allows sensitive analysis of glycans benefiting from optimized deglycosylation reactions and efficient sample cleanup. Copyright © 2005 John Wiley & Sons, Ltd.