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Fully automated two-step assay for detection of metallothionein through magnetic isolation using functionalized γ-Fe2O3 particles
MA. Merlos Rodrigo, L. Krejcova, J. Kudr, N. Cernei, P. Kopel, L. Richtera, A. Moulick, D. Hynek, V. Adam, M. Stiborova, T. Eckschlager, Z. Heger, O. Zitka,
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.
2016 ;
1039 (-) :
17-27.
[pub] 20161014
Language English Country Netherlands
Document type Journal Article
Persistent link
https://www.medvik.cz/link/bmc17013415
Grant support
NV15-28334A
MZ0
CEP Register
- MeSH
- Electrophoresis, Polyacrylamide Gel methods MeSH
- Chromatography, Gel methods MeSH
- Liver chemistry MeSH
- Rabbits MeSH
- Rats MeSH
- Automation, Laboratory MeSH
- Magnetite Nanoparticles chemistry MeSH
- Metallothionein analysis isolation & purification MeSH
- Rats, Wistar MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Rats MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
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.
References provided by Crossref.org
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