title: Online LC-FAIMS-MS/MS for the Analysis of Phosphorylation in Proteins.
abstract: High-field asymmetric waveform ion mobility spectrometry (FAIMS) is a gas-phase separation technique which, when coupled with liquid chromatography tandem mass spectrometry, offers benefits for analysis of complex proteomics samples such as those encountered in phosphoproteomics experiments. Results from LC-FAIMS-MS/MS are typically complementary, in terms of proteome coverage and isomer identification, to those obtained by use of solution-phase separation methods, such as prefractionation with strong cation-exchange chromatography. Here, we describe the protocol for large-scale phosphorylation analysis by LC-FAIMS-MS/MS.
authors.affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Zhao
authors.affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Creese
authors.affiliation: School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK. h.j.cooper@bham.ac.uk.
authors.forename: Helen J
authors.initials: HJ
authors.lastname: Cooper
mesh_terms: D000818:Animals; D056148:Chromatography, Reverse-Phase / Q000295:instrumentation; D019295:Computational Biology; D030562:Databases, Protein; D006801:Humans; D009862:Online Systems; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000295:instrumentation; D057188:Workflow
chemical_list: D010750:Phosphoproteins
keywords: FAIMS; Ion mobility spectrometry; Phosphorylation