Characterization of pericardial fluid proteome through fractionation with magnetic nanoparticles (NPs@EDTA)

ABSTRACT: Despite the invasive nature of its collection, being in close contact to the heart, the pericardial fluid (PF) proteome reflects the pathophysiological status of this organ. Therefore, whenever it is available, e.g. on occasion of open-heart surgery, PF proteomic analysis can give important clues of the molecular mechanisms underlying heart disease and it may help uncover new diagnostic/prognostic markers as well as new therapeutic targets. Still, the characterization of its protein fraction is challenged by large amounts of albumin, that can be relatively higher than in plasma/serum. To help expand the PF catalogued to date and to validate the technique for future applications, we have fractionated and characterized the pericardial fluid, using N-(trimethoxysilylpropyl)ethylenediamine triacetic acid (EDTA)-functionalized magnetic nanoparticles (NPs@EDTA) followed by a GeLC-MS/MS approach. As compared to a traditional albumin and IgG-depletion kit, we found this method to be as efficient in removing albumin, with similar inter-individual proteome variability. Furthermore, the NPs@EDTA-based fractionation approach could expand the protein dynamic range and displayed good proteome reproducibility. Overall, 565 proteins were identified, with more than half being new assignments to PF proteome.

INSTRUMENT(S): LTQ Orbitrap Velos, TripleTOF 5600

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Pericardial Fluid

SUBMITTER: Fábio Trindade  

LAB HEAD: Rui Miguel Pinheiro Vitorino

PROVIDER: PXD008620 | Pride | 2018-01-09

REPOSITORIES: Pride