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Capillary zone electrophoresis coupled to drift tube ion mobility-mass spectrometry for the analysis of native and APTS-labeled N-glycans

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Abstract

Capillary zone electrophoresis (CZE) based on electrophoretic mobility in the liquid phase and ion mobility spectrometry (IMS) based on mobilities in the gas phase are both powerful techniques for the separation of complex samples. Protein glycosylation is one of the most common post-translational modifications associated with a wide range of biological functions and human diseases. Due to their high structural variability, the analysis of glycans still represents a challenging task. In this work, the first on-line coupling of CZE with drift tube ion mobility-mass spectrometry (DTIM-MS) has been perfomed to further improve separation capabilities for the analysis of native and 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled N-glycans. In this way, a complexity of glycan signals was revealed which could not be resolved by these techniques individually, shown for both native and APTS-labeled glycans. Each individual glycan signal separated in CZE exhibited an unexpectedly high number of peaks observed in the IMS dimension. This observation could potentially be explained by the presence of isomeric forms, including different linkages, and/or gas-phase conformers. In addition, the type of sialic acid attached to glycans has a significant impact on the obtained drift time profile. Furthermore, the application of α2-3 neuraminidase enabled the partial assignment of peaks in the arrival time distribution considering their sialic acid linkages (α2-3/α2-6). This work is a showcase for the high potential of CZE-DTIM-MS, which is expected to find various applications in the future.

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Abbreviations

AGP:

α-1-Acid glycoprotein

APTS:

8-Aminopyrene-1,3,6-trisulfonic acid

ATD:

Arrival time distribution

BGE:

Background electrolyte

CE:

Capillary electrophoresis

CZE:

Capillary zone electrophoresis

DT:

Drift time

DTIM-MS:

Drift tube ion mobility-mass spectrometry

ESI:

Electrospray ionization

FAIMS-MS:

High-field asymmetric waveform ion mobility spectrometry mass spectrometry

HILIC:

Hydrophilic interaction chromatography

IMS:

Ion mobility spectrometry

mAb:

Monoclonal antibody

MS:

Mass spectrometry

NeuNAc:

N-Acetylneuraminic acid

NeuNGc:

N-Glycolylneuraminic acid

SA:

Sialic acid

SL:

Sheath liquid

TOF-MS:

Time-of-flight mass spectrometry

TWIM-MS:

Traveling wave ion mobility-mass spectrometry

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Authors and Affiliations

  1. Faculty of Chemistry, Aalen University, 73430, Aalen, Germany

    Kevin Jooß & Christian Neusüß

  2. Research Unit Analytical BioGeoChemistry, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany

    Kevin Jooß

  3. Applied Analytical Chemistry, University of Duisburg-Essen, Universitätsstr. 5, 45141, Essen, Germany

    Sven W. Meckelmann, Julia Klein & Oliver J. Schmitz

  4. Teaching and Research Center for Separation, University of Duisburg-Essen, Universitätsstr. 5, 45141, Essen, Germany

    Sven W. Meckelmann, Julia Klein & Oliver J. Schmitz

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  1. Kevin Jooß
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  2. Sven W. Meckelmann
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Correspondence to Christian Neusüß.

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Published in the topical collection Close-Up of Current Developments in Ion Mobility Spectrometry with guest editor Gérard Hopfgartner.

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Jooß, K., Meckelmann, S.W., Klein, J. et al. Capillary zone electrophoresis coupled to drift tube ion mobility-mass spectrometry for the analysis of native and APTS-labeled N-glycans. Anal Bioanal Chem 411, 6255–6264 (2019). https://doi.org/10.1007/s00216-018-1515-7

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  • DOI: https://doi.org/10.1007/s00216-018-1515-7

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