title: 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.
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.
authors.affiliation: CICECO, Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal.
authors.forename: Cláudia
authors.initials: C
authors.lastname: Couto
authors.affiliation: QOPNA, University of Aveiro, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal; Faculty of Pharmacy and Centre for Neuroscience and Cell Biology, University of Coimbra 3000-548, Coimbra - Portugal.
authors.forename: Bruno
authors.initials: B
authors.lastname: Neves
authors.affiliation: QOPNA, University of Aveiro, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal.
authors.forename: Rita
authors.initials: R
authors.lastname: Ferreira
authors.affiliation: CICECO, Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal. Electronic address: ana.luisa@ua.pt.
authors.forename: Ana L
authors.initials: AL
authors.lastname: Daniel-da-Silva
authors.affiliation: Department of Medical Sciences, Institute for Biomedicine - iBiMED, University of Aveiro, Aveiro, Portugal; Department of Physiology and Cardiothoracic Surgery, Faculty of Medicine, University of Porto, Porto, Portugal. Electronic address: rvitorino@ua.pt.
authors.forename: Rui
authors.initials: R
authors.lastname: Vitorino
mesh_terms: D000085:Acetates / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D002614:Chelating Agents / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods; D005029:Ethylenediamines / Q000737:chemistry; D006801:Humans; D008264:Macrophages / Q000737:chemistry*; D060328:Magnetic Phenomena; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D051379:Mice; D009000:Monocytes / Q000737:chemistry*; D009211:Myoglobin / Q000032:analysis* / Q000737:chemistry; D017646:Organosilicon Compounds / Q000737:chemistry; D020543:Proteome / Q000032:analysis; D040901:Proteomics; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry; D014357:Trypsin / Q000032:analysis* / Q000737:chemistry
chemical_list: D000085:Acetates; D002614:Chelating Agents; D005029:Ethylenediamines; D058185:Magnetite Nanoparticles; D009211:Myoglobin; D017646:Organosilicon Compounds; D020543:Proteome; D012710:Serum Albumin, Bovine; D014357:Trypsin
keywords: Chelating chromatography; Magnetic nanoparticles; Mass spectrometry; Metalloproteins; Proteomics; Secretome; albumin depletion