Fully automated two-step assay for detection of metallothionein through magnetic isolation using functionalized γ-Fe2O3 particles.

Abstract

Metallothioneins (MTs) are involved in heavy metal detoxification in a wide range of living organisms. Currently, it is well known that 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 cancer diagnostics, an easy-to-use and rapid method for its detection is required. Hence, the aim of this study was to develop a fully automated and high-throughput assay for the estimation of MT levels. Here, we report the optimal conditions for the isolation of MTs from rabbit liver and their characterization using MALDI-TOF MS. In addition, we described a two-step assay, which started with an isolation of the protein using functionalized paramagnetic particles and finished with their electrochemical analysis. The designed easy-to-use, cost-effective, error-free and fully automated procedure for the isolation of MT coupled with a simple analytical detection method can provide a prototype for the construction of a diagnostic instrument, which would be appropriate for the monitoring of carcinogenesis or MT-related chemoresistance of tumors.

• Authors: Miguel Angel Merlos Rodrigo, Ludmila Krejcova, Jiri Kudr, Natalia Cernei, Pavel Kopel, Lukas Richtera, Amitava Moulick, David Hynek, Vojtech Adam, Marie Stiborova, Tomas Eckschlager, Zbynek Heger, Ondrej Zitka
• Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (J Chromatogr B Analyt Technol Biomed Life Sci) Vol. 1039 Pg. 17-27 (Dec 15 2016) ISSN: 1873-376X [Electronic] Netherlands
• PMID: 27825623 (Publication Type: Journal Article)
• Copyright: Copyright © 2016 Elsevier B.V. All rights reserved.

Chemical References

• Magnetite Nanoparticles
• Metallothionein

Topics

• Animals
• Automation, Laboratory
• Chromatography, Gel (methods)
• Electrophoresis, Polyacrylamide Gel (methods)
• Liver (chemistry)
• Magnetite Nanoparticles (chemistry)
• Male
• Metallothionein (analysis, isolation & purification)
• Rabbits
• Rats
• Rats, Wistar
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (methods)