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 |
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| Rok vydání: | 2017 |
| Předmět: |
Electrophoresis
0301 basic medicine Proteome Biophysics Fractionation Chemical Fractionation Proteomics Biochemistry Mass Spectrometry 03 medical and health sciences Humans Chelation Magnetite Nanoparticles Molecular Biology Chelating Agents Chromatography 030102 biochemistry & molecular biology Chemistry Albumin Pericardial fluid Binding ability 030104 developmental biology Pericardial Fluid Magnetic nanoparticles Adsorption |
| 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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