Precursor acquisition independent from ion count: how to dive deeper into the proteomics ocean.
Data-dependent precursor ion selection is widely used in shotgun proteomics to profile the protein components of complex samples. Although very popular, this bottom-up method presents major drawbacks in terms of detectable dynamic range. Here, we demonstrate the superior performance of a data-independent method we term precursor acquisition independent from ion count (PAcIFIC). Our results show that almost the entire, predicted, soluble bacterial proteome can be thoroughly analyzed by PAcIFIC without the need for any sample fractionation other than the C18-based liquid chromatograph used to introduce the peptide mixture into the mass spectrometer. Importantly, we also show that PAcIFIC provides unique performance for analysis of human plasma in terms of the number of proteins identified (746 at FDR < or = 0.5%) and achieved dynamic range (8 orders of magnitude at FDR < or = 0.5%), without any fractionation other than immuno-depletion of the seven most abundant proteins.
Panchaud
Alexandre
A
Department of Medicinal Chemistry, University of Washington, Seattle, Washington 98195, USA.
Scherl
Alexander
A
Shaffer
Scott A
SA
von Haller
Priska D
PD
Kulasekara
Hemantha D
HD
Miller
Samuel I
SI
Goodlett
David R
DR
D001426:Bacterial Proteins / Q000032:analysis*; D001798:Blood Proteins / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D019295:Computational Biology; D006801:Humans; D007477:Ions / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D011550:Pseudomonas aeruginosa / Q000254:growth & development / Q000378:metabolism; D053719:Tandem Mass Spectrometry
D001426:Bacterial Proteins; D001798:Blood Proteins; D007477:Ions; D010446:Peptide Fragments; D020543:Proteome