Contribution of Capillary Zone Electrophoresis Hyphenated with Drift Tube Ion Mobility Mass Spectrometry as a Complementary Tool to Microfluidic Reversed Phase Liquid Chromatography for Antigen Discovery

Marie-Jia Gou; Murat Cem Kose; Jacques Crommen; Cindy Nix; Gael Cobraiville; Jo Caers; Marianne Fillet

doi:10.3390/ijms232113350

Contribution of Capillary Zone Electrophoresis Hyphenated with Drift Tube Ion Mobility Mass Spectrometry as a Complementary Tool to Microfluidic Reversed Phase Liquid Chromatography for Antigen Discovery

Int J Mol Sci. 2022 Nov 1;23(21):13350. doi: 10.3390/ijms232113350.

Authors

Marie-Jia Gou¹, Murat Cem Kose², Jacques Crommen¹, Cindy Nix¹, Gael Cobraiville¹, Jo Caers^{2

3}, Marianne Fillet¹

Affiliations

¹ Laboratory for the Analysis of Medicines (LAM), Department of Pharmacy, CIRM, University of Liege, Avenue Hippocrate 15, B36 Tour 4 +3, 4000 Liège, Belgium.
² Laboratory of Hematology, GIGA I3, University of Liège, Avenue de l'Hopital 11, B34, 4000 Liege, Belgium.
³ Department of Hematology, Centre Hospitalier Universitaire (CHU) de Liège, Avenue de l'Hopital 1, 4000 Liege, Belgium.

Abstract

The discovery of new antigens specific to multiple myeloma that could be targeted by novel immunotherapeutic approaches is currently of great interest. To this end, it is important to increase the number of proteins identified in the sample by combining different separation strategies. A capillary zone electrophoresis (CZE) method, coupled with drift tube ion mobility (DTIMS) and quadrupole time-of-flight mass spectrometry (QTOF), was developed for antigen discovery using the human myeloma cell line LP-1. This method was first optimized to obtain a maximum number of identifications. Then, its performance in terms of uniqueness of identifications was compared to data acquired by a microfluidic reverse phase liquid chromatography (RPLC) method. The orthogonality of these two approaches and the physicochemical properties of the entities identified by CZE and RPLC were evaluated. In addition, the contribution of DTIMS to CZE was investigated in terms of orthogonality as well as the ability to provide unique information. In conclusion, we believe that the combination of CZE-DTIMS-QTOF and microfluidic RPLC provides unique information in the context of antigen discovery.

Keywords: capillary zone electrophoresis mass spectrometry; drift tube ion mobility spectrometry; orthogonality; untargeted proteomics.

MeSH terms

Cell Line, Tumor
Chromatography, Reverse-Phase*
Electrophoresis, Capillary / methods
Humans
Microfluidics
Multiple Myeloma*
Tandem Mass Spectrometry / methods

Grants and funding

M.-J.G. is an FRIA grantee of the Fonds de la Recherche Scientifique–FNRS and also received funding from the Fondation Léon Frédéricq (Grant number: 2021-2022–60). The authors acknowledge the research grants from the Walloon Region of Belgium and the EU Commission (project FEDER-PHARE) for the funding of the DTIMS–QTOF instrument.