Enhanced sensitivity in proteomics experiments using FAIMS coupled with a hybrid linear ion trap/Orbitrap mass spectrometer.
We describe the use and application of high-field asymmetric waveform ion mobility spectrometry combined with nanoscale liquid chromatography mass spectrometry (nanoLC-FAIMS-MS) to improve the sensitivity and dynamic range of proteomics analyses on a hybrid LTQ-Orbitrap mass spectrometer. The ability of FAIMS to enrich multiply protonated peptides against background ions confers a marked advantage in proteomics analyses by decreasing the limits of detection to facilitate the identification of low-abundance peptide ions. These multiply charged ions are recorded into separate acquisition channels to enhance the overall population of detectable peptide ions from a single analysis. NanoLC-FAIMS-MS experiments performed on peptides spiked into complex proteins digests provided more than 10-fold improvement in limits of detection compared to conventional nanoelectrospray mass spectrometry. This enhancement of sensitivity is reflected by a 55% increase in the number of assigned MS/MS spectra contributing to an overall improvement in protein identification and sequence coverage. The application of FAIMS in label-free quantitative proteomics is demonstrated for the identification of differentially abundant proteins from human U937 monocytic cells exposed to phorbol ester.
Saba
Julian
J
Institute for Research in Immunology and Cancer, Université de Montréal, Department of Chemistry, P.O. Box 6128, Station Centre-ville, Montréal, Canada H3C 3J7.
Bonneil
Eric
E
Pomiès
Christelle
C
Eng
Kevin
K
Thibault
Pierre
P
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D045304:Cytochromes c / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D009000:Monocytes / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D011817:Rabbits; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D020298:U937 Cells
D010455:Peptides; D020543:Proteome; D045304:Cytochromes c