title: Increased throughput of proteomics analysis by multiplexing high-resolution tandem mass spectra.
abstract: High-resolution and high-accuracy Fourier transform mass spectrometry (FTMS) is becoming increasingly attractive due to its specificity. However, the speed of tandem FTMS analysis severely limits the competitive advantage of this approach relative to faster low-resolution quadrupole ion trap MS/MS instruments. Here we demonstrate an entirely FTMS-based analysis method with a 2.5-3.0-fold greater throughput than a conventional FT MS/MS approach. The method consists of accumulating together the MS/MS fragments ions from multiple precursors, with subsequent high-resolution analysis of the mixture. Following acquisition, the multiplexed spectrum is deconvoluted into individual MS/MS spectra which are then combined into a single concatenated file and submitted for peptide identification to a search engine. The method is tested both in silico using a database of MS/MS spectra as well as in situ using a modified LTQ Orbitrap mass spectrometer. The performance of the method in the experiment was consistent with theoretical expectations.
authors.affiliation: Department of Chemistry and Biomolecular Chemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706-1322, USA.
authors.forename: A R
authors.initials: AR
authors.lastname: Ledvina
authors.forename: M M
authors.initials: MM
authors.lastname: Savitski
authors.forename: A R
authors.initials: AR
authors.lastname: Zubarev
authors.forename: D M
authors.initials: DM
authors.lastname: Good
authors.forename: J J
authors.initials: JJ
authors.lastname: Coon
authors.forename: R A
authors.initials: RA
authors.lastname: Zubarev
mesh_terms: D002851:Chromatography, High Pressure Liquid; D005583:Fourier Analysis; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D010455:Peptides; D011506:Proteins