title: Balancing robust quantification and identification for iTRAQ: application of UHR-ToF MS.
abstract: iTRAQ reagents allow the simultaneous multiplex identification and quantification of a large number of proteins. Success depends on effective peptide fragmentation in order to generate both peptide sequence ions (higher mass region, 150-2200 m/z) and reporter ions (low mass region, 113-121 m/z) for protein identification and relative quantification, respectively. After collision-induced dissociation, the key requirements to achieve a good balance between the high and low m/z ions are effective ion transmission and detection across the MS/MS mass range, since the ion transmission of the higher m/z range competes with that of the low m/z range. This study describes an analytical strategy for the implementation of iTRAQ on maXis UHR-Qq-ToF instruments, and discusses the impact of adjusting the MS/MS ion transmission parameters on the quality of the overall data sets. A technical discussion highlights a number of maXis-specific parameters, their impact of quantification and identification, and their cross-interactions.
authors.affiliation: ChELSI Institute, Department of Chemical and Process Engineering, University of Sheffield, Sheffield, UK.
authors.forename: Saw Yen
authors.initials: SY
authors.lastname: Ow
authors.forename: Josselin
authors.initials: J
authors.lastname: Noirel
authors.forename: Malinda
authors.initials: M
authors.lastname: Salim
authors.forename: Caroline
authors.initials: C
authors.lastname: Evans
authors.forename: Rod
authors.initials: R
authors.lastname: Watson
authors.forename: Phillip C
authors.initials: PC
authors.lastname: Wright
mesh_terms: D013058:Mass Spectrometry / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry