Currently, it is well known that metallothioneins (MTs) play substantial role in many pathophysiological processes, including carcinogenesis and they can serve as diagnostic biomarkers. In order to increase the applicability of MT in diagnostics, an easy-to-use and rapid detection method is required. The aim study was to develop a fully automated and high-throughput assay for the estimation of MT levels. We focused on the design and fabrication of a method based on isolation of MTs using paramagnetic particles (functionalized nanomaghemite γ-Fe2O3 core) with consequent detection by differential pulse voltammetry (DPV) utilizing Brdicka electrolyte. We designed and tested six types of particles, which differed in the surface functionalization [polyvinylpyrrolidone and HAuCl4; polyethyleneimine; polytheleneglycol (MW 1500 and 4000) and HAuCl4; poly(4-styrenesulfonic acid and HAuCl4; and polyacrylamide and HAuCl4]. The best conditions were employed in the automation of isolation and detection procedure, which made it simple and fast. As a proof-of-concept we successfully applied our protocol for isolation and detection of MT in serum collected from Wistar rats. The designed easy-to-use, cost-effective and fully automated procedure for the isolation of MT coupled with a simple electrochemical detection can serve for the construction of a diagnostic instrument, which would be appropriate for the monitoring of carcinogenesis or MT-related chemoresistance of tumors.Keywords: Automation; electrochemistry; magnetic isolation; MALDI-TOF MS; metallothionein
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