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Comparative ESI FT-MS and MALDI-TOF structural analyses of representative human N-linked glycans

Zuzana Pakanová, Marek Nemčovič, Peter Bystrický, Mária Matulová, Vladimír Pätoprstý, Iain B. H. Wilson, and Ján Mucha

Institute of Chemistry, Center of Excellence for Glycomics, Slovak Academy of Sciences, Dúbravská cesta 9, 84538 Bratislava, Slovakia

 

E-mail: zuzana.pakanova@savba.sk

Abstract: Modern glycan analysis is primarily based on mass spectrometry, where instruments based on electrospray or matrix-assisted laser desorption ionization are currently the most frequently used. In the present study, electrospray ionization (ESI) coupled with a high-resolution Fourier transform mass spectrometer (LTQ Orbitrap) and matrix-assisted laser desorption/ionization (MALDI) coupled with a time-of-flight (TOF/TOF) detector were used to analyze two N-glycan standards with intact free reducing ends (disialo biantennary and asialo triantennary) and representative PA-labeled human serum N-glycan structures isolated by hydrophilic interaction anion-exchange chromatography (HIAX), confirmed by 1H NMR analysis and consequently compared with the ProteinScape Glycome database. Different combinations of ion sources with fragmentation devices results in various fragmentation patterns and adducts. Also, the effect of sample derivatization on the acquired signals is discussed. Compared to the MALDI technique, free glycans did not lose labile sialic acids easily in the ESI source. On the other hand, fluorescent PA-labeling leads to improved core fragmentation and signal intensities; linkage-specific ethyl esterification leads to reduced adduct and fragment formation and enhanced stability of sialic acids in the MALDI ion source. Thereby, both methods have their advantages and disadvantages in terms of detection, fragmentation and robustness.

Keywords: N-glycans – glycosylation – MALDI – ESI – glycoprofiling

Full paper is available at www.springerlink.com.

DOI: 10.1515/chempap-2015-0182

 

Chemical Papers 69 (12) 1633–1638 (2015)

Sunday, November 28, 2021

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