Imaging mass spectrometry using peptide isoelectric focusing |
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Authors: | Vaezzadeh Ali R Simicevic Jovan Chauvet Alexis François Patrice Zimmermann-Ivol Catherine G Lescuyer Pierre Deshusses Jacques P M Hochstrasser Denis F |
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Affiliation: | Biomedical Proteomics Research Group, Department of Structural Biology and Bioinformatics, Geneva University, 1 rue Michel Servet, 1211 Geneva, Switzerland. alireza.vaezzadeh@medecine.unige.ch |
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Abstract: | Imaging Mass Spectrometry (IMS) has emerged as a powerful technique in the field of proteomics. The use of Immobilized pH Gradient-IsoElectric Focusing (IPG-IEF) is also a new trend, as the first dimension of separation, in shotgun proteomics. We report a combination of these two outstanding technologies. This approach is based on the separation of shotgun-produced peptides by IPG-IEF. The peptides are then transferred by capillarity to a capture membrane, which is then scanned by the mass spectrometer to generate MS images. This high-throughput methodology allows a preview of the sample to be obtained in a single day. We report the application of this new pipeline for differential comparison of the membrane proteome of two different strains of Staphylococcus aureus bacteria in a proof-of-principle experiment. |
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