Improvement in Peptide Detection for Proteomics Analyses Using NanoLC-MS and High-Field Asymmetry Waveform Ion Mobility Mass Spectrometry

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• 作者：Karine Venne ; Eric Bonneil ; Kevin Eng ; and Pierre Thibault
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：April 1, 2005
• 年：2005
• 卷：77
• 期：7
• 页码：2176 - 2186
• 全文大小：663K
• 年卷期：v.77,no.7(April 1, 2005)
• ISSN：1520-6882

文摘

Sensitive and selective detection of multiply chargedpeptide ions from complex tryptic digests was achievedusing high-field asymmetric waveform ion mobility spectrometry (FAIMS) combined with nanoscale liquid chromatography-mass spectrometry (nanoLC-FAIMS-MS).The combination of FAIMS provided a marked advantageover conventional nanoLC-MS experiments by reducingthe extent of chemical noise associated with singly chargedions and enhancing the overall population of detectabletryptic peptides. Such advantages were evidenced by a6-12-fold improvement in signal-to-noise ratio measurements for a wide range of multiply charged peptide ions.An increase of 20% in the number of detected peptidescompared to conventional nanoelectrospray was achievedby transmitting ions of different mobilities at high electricfield vs low field while simultaneously recording each ionpopulation in separate mass spectrometry acquisitionchannels. This method provided excellent reproducibilityacross replicate nanoLC-FAIMS-MS runs with more than90% of all detected peptide ions showing less than 30%variation in intensity. The application of this techniquein the context of proteomics research is demonstrated forthe identification of trace-level proteins showing differential expression in U937 monocyte cell extracts following incubation with phorbol ester.