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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Keywords
Glycan analysis CE-IM-MS Isomer separation Liquid- and gas-phase separationAbbreviations
- 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
Notes
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Supplementary material
References
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