A fractionation approach applying chelating magnetic nanoparticles to characterize pericardial fluid's proteome

Autor: Sofia F. Soares, Paulo Bastos, Rita Ferreira, Adelino F. Leite-Moreira, Inês Falcão-Pires, Ana L. Daniel-da-Silva, Rui Vitorino, Fábio Trindade, Bruno Manadas
Rok vydání: 2017
Předmět:
Zdroj: Archives of Biochemistry and Biophysics. 634:1-10
ISSN: 0003-9861
DOI: 10.1016/j.abb.2017.09.016
Popis: Owing to their close proximity, pericardial fluid (PF)'s proteome may mirror the pathophysiological status of the heart. Despite this diagnosis potential, the knowledge of PF's proteome is scarce. Large amounts of albumin hamper the characterization of the least abundant proteins in PF. Aiming to expand PF's proteome and to validate the technique for future applications, we have fractionated and characterized the PF, using N-(trimethoxysilylpropyl)ethylenediamine triacetic acid (EDTA)-functionalized magnetic nanoparticles (NPs@EDTA) followed by a GeLC-MS/MS approach. Similarly to an albumin-depletion kit, NPs@EDTA-based fractionation was efficient in removing albumin. Both methods displayed comparable inter-individual variability, but NPs@EDTA outperformed the former with regard to the protein dynamic range as well as to the monitoring of biological processes. Overall, 565 proteins were identified, of which 297 (>50%) have never been assigned to PF. Moreover, owing to this method's good proteome reproducibility, affordability, rapid automation and high binding ability of NP@EDTA, it bears a great potential towards future clinical application.
Databáze: OpenAIRE
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