Scanning quadrupole DIA - Scanning Quadrupole Data Independent Acquisition - Part A

ABSTRACT: A scanning quadrupole data independent acquisition (DIA) method is described and characterised for the qualitative and quantitative label free proteomic analysis of complex biological samples. The principle of the scanning quadrupole DIA is discussed and the effect of the optimization of analytical instrument characteristics, such as acquisition speed, sample complexity, and scan/integration time in relation to sample complexity for a number of different model proteomes of various complexity and dynamic range, including plasma, human cell lines, and bacteria samples, discussed. In addition, its technological merits from an acquisition neutral perspective are reviewed. The qualitative and semi-quantitative performance of the method is illustrated and contrasted for the analysis of a well-characterised human cell line sample using untargeted and targeted search strategies. The application of the method is demonstrated for the analysis of the calcineurin-dependent proteome of drug treated Aspergillus fumigatus. The design of these experiments afforded assessment of the precision and accuracy of scanning quadrupole data independent acquisition (DIA) method as will be demonstrated by peptide and protein centric analysis of the study pool QC samples of the study. Moreover, novel and specific interactors in response to drug treatment that are indicative of the role of calcineurin role in regulating these effectors have been identified.

INSTRUMENT(S): Xevo G2-S QTof

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Hans Vissers  

LAB HEAD: JPC Vissers

PROVIDER: PXD005869 | Pride | 2017-09-14

REPOSITORIES: pride