Enhanced Sensitivity in Proteomics Experiments Using FAIMS Coupled with a Hybrid Linear Ion Trap/Orbitrap Mass Spectrometer

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• 作者：Julian Saba ; Eric Bonneil ; Christelle Pomis ; Kevin Eng ; Pierre Thibault
• 刊名：Journal of Proteome Research
• 出版年：2009
• 出版时间：July 6, 2009
• 年：2009
• 卷：8
• 期：7
• 页码：3355-3366
• 全文大小：414K
• 年卷期：v.8,no.7(July 6, 2009)
• ISSN：1535-3907

文摘

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.