title: Automated Validation of Results and Removal of Fragment Ion Interferences in Targeted Analysis of Data-independent Acquisition Mass Spectrometry (MS) using SWATHProphet.
abstract: Proteomics by mass spectrometry technology is widely used for identifying and quantifying peptides and proteins. The breadth and sensitivity of peptide detection have been advanced by the advent of data-independent acquisition mass spectrometry. Analysis of such data, however, is challenging due to the complexity of fragment ion spectra that have contributions from multiple co-eluting precursor ions. We present SWATHProphet software that identifies and quantifies peptide fragment ion traces in data-independent acquisition data, provides accurate probabilities to ensure results are correct, and automatically detects and removes contributions to quantitation originating from interfering precursor ions. Integration in the widely used open source Trans-Proteomic Pipeline facilitates subsequent analyses such as combining results of multiple data sets together for improved discrimination using iProphet and inferring sample proteins using ProteinProphet. This novel development should greatly help make data-independent acquisition mass spectrometry accessible to large numbers of users.
authors.affiliation: From the Institute for Systems Biology, Seattle, Washington 98109.
authors.forename: Andrew
authors.initials: A
authors.lastname: Keller
authors.affiliation: From the Institute for Systems Biology, Seattle, Washington 98109.
authors.forename: Samuel L
authors.initials: SL
authors.lastname: Bader
authors.affiliation: From the Institute for Systems Biology, Seattle, Washington 98109.
authors.forename: David
authors.initials: D
authors.lastname: Shteynberg
authors.affiliation: From the Institute for Systems Biology, Seattle, Washington 98109.
authors.forename: Leroy
authors.initials: L
authors.lastname: Hood
authors.affiliation: From the Institute for Systems Biology, Seattle, Washington 98109 robert.moritz@systemsbiology.org.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Moritz
mesh_terms: D006801:Humans; D019151:Peptide Library; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D011507:Proteinuria / Q000652:urine*; D059748:Proteolysis; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D012984:Software; D053719:Tandem Mass Spectrometry / Q000706:statistics & numerical data*; D014357:Trypsin / Q000737:chemistry
chemical_list: D019151:Peptide Library; D010455:Peptides; D011506:Proteins; D014357:Trypsin