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Sensors 2008, 8(4), 2293-2305; doi:10.3390/s8042293
Article

An Electrochemical Detection of Metallothioneins at the Zeptomole Level in Nanolitre Volumes

1,2, 1, 1,3, 4 and 1,*
1 Department of Chemistry and Biochemistry Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic 2 Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic 3 Department of Chemistry Faculty of Science, Masaryk University, Kotlarska 2, CZ-611 37 Brno, Czech Republic 4 Department of Chemistry, Faculty of Science, Masaryk University, Kotlarska 2, CZ-611 37 Brno, Czech Republic
* Author to whom correspondence should be addressed.
Received: 15 March 2008 / Accepted: 26 March 2008 / Published: 1 April 2008
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Abstract

An Electrochemical Detection of Metallothioneins at the Zeptomole Level in Nanolitre VolumesWe report on improvement of the adsorptive transfer stripping technique (AdTS) coupled with the differential pulse voltammetry Brdicka reaction to determine a thiol-protein. The current technique has been unable to generate reproducible results when analyzing very low sample volumes (nanolitres). This obstacle can be overcome technically by modifying the current transfer technique including cooling step of the adsorbed analyte. We tested the technique on determination of a promising tumour disease marker protein called metallothionein (MT). The detection limit (3 S/N) of MT was evaluated as 500 zeptomoles per 500 nL (1 pM) and the quantification limit (10 S/N) as 1,500 zeptomoles per 500 nL (3 pM). Further, the improved AdTS technique was utilized to analyze blood serum samples from patients with breast cancer. Based on the results obtained it can be concluded that the improved technique can be used to detect a thiolprotein in very low sample volumes and can also prevent interferences during the washing and transferring step.
Keywords: Proteomics; metallothionein; thiols; differential pulse voltammetry; Brdicka reaction; adsorptive transfer stripping technique; human blood serum; tumour disease; zeptomole Proteomics; metallothionein; thiols; differential pulse voltammetry; Brdicka reaction; adsorptive transfer stripping technique; human blood serum; tumour disease; zeptomole
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Adam, V.; Baloun, J.; Fabrik, I.; Trnkova, L.; Kizek, R. An Electrochemical Detection of Metallothioneins at the Zeptomole Level in Nanolitre Volumes. Sensors 2008, 8, 2293-2305.

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