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Reputable Mentor II
Reputable Mentor II
Hoopmann MR, Merrihew GE, von Haller PD, MacCoss MJ.
J Proteome Res. 2009 Apr;8(4):1870-5.
The identification of peptides by microcapillary liquid chromatography-tandem mass spectrometry (microLC-MS/MS) has become routine because of the development of fast scanning mass spectrometers, data-dependent acquisition, and database searching algorithms. However, many peptides within the detection limit of the mass spectrometer remain unidentified because of limitations in MS/MS sampling speed despite the dynamic range and peak capacity of the instrument. We have developed an automated approach that uses the mass spectra from high resolution microLC-MS data to define the molecular species present in the mixture and directs the acquisition of MS/MS spectra to precursors that were missed in prior analyses. This approach increases the coverage of the molecular species sampled by MS/MS and consequently the number of peptides and proteins identified during the acquisition of technical or biological replicates using a simple one-dimensional chromatographic separation. The combination of a unique workflow and custom software contribute to the improved identification of molecular features detected in proteomics experiments of complex protein mixtures.
University of Washington, Department of Genome Sciences, Seattle, Washington 98195, USA.
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