Proteomic studies with a novel nano-magnetic chelating system to capture metalloproteins and its application in the preliminary study of monocyte and macrophage sub-secretome
authors Couto, C; Neves, B; Ferreira, R; Daniel-da-Silva, AL; Vitorino, R
nationality International
journal TALANTA
author keywords Magnetic nanoparticles; Metalloproteins; Secretome; Chelating chromatography; Mass spectrometry; Proteomics; albumin depletion
keywords PROTEIN; TOOLS; CELLS
abstract A new chelating chromatography method was developed based in the use of magnetic iron oxide nanoparticles functionalized with EDTA-TMS ((N-(trimethoxysilylpropyl)ethylenediaminetriacetate trisodium salt). These particles combine a high surface area, biocompatibility and magnetic removal from solution, with the chelating affinity towards metal ions. The particles were used to selectively capture metallo-dependant proteins in secretome obtained from human monocytes and mouse macrophages. Secreted metallo-dependant proteins are highly important sources of information since they are involved in several pathological processes. The identification of secreted proteins involved in these processes is highly important for diagnosis or monitoring the progression of a disease. In this multiple-approach study it was possible to not only selectively capture several secreted metallo-dependant proteins, but also to significantly avoid masking proteins such as the highly abundant albumin form the fetal bovine serum used to supplement the cell culture medium. Overall, the magnetic nanoparticle-based chelating chromatography method developed here has proved to be a sensitive, low cost, and a quick tool for sample treatment in order to selectively enrich metalloproteins while overcoming the contamination of highly abundant proteins. (C) 2016 Elsevier B.V. All rights reserved.
publisher ELSEVIER SCIENCE BV
issn 0039-9140
year published 2016
volume 158
beginning page 110
ending page 117
digital object identifier (doi) 10.1016/j.talanta.2016.05.051
web of science category Chemistry, Analytical
subject category Chemistry
unique article identifier WOS:000379370400015
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journal impact factor 5.339
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