Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 1039, Issue -, Pages 17-27Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2016.10.018
Keywords
Automation; Biomarkers; Magnetic isolation; MALDI-TOF MS; Metallothionein
Funding
- Ministry of Education, Youth and Sports of the Czech Republic [LO1401]
- Ministry of Health of the Czech Republic [00064203]
- [MTCYTO15-28334A]
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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. (C) 2016 Elsevier B.V. All rights reserved.
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