title: An Electrochemical Detection of Metallothioneins at the Zeptomole Level in Nanolitre Volumes.
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.
authors.affiliation: Department of Chemistry and Biochemistry Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic.|Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic.
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
authors.affiliation: Department of Chemistry and Biochemistry Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic.
authors.forename: Jiri
authors.initials: J
authors.lastname: Baloun
authors.affiliation: Department of Chemistry and Biochemistry Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic.|Department of Chemistry Faculty of Science, Masaryk University, Kotlarska 2, CZ-611 37 Brno, Czech Republic.
authors.forename: Ivo
authors.initials: I
authors.lastname: Fabrik
authors.affiliation: Department of Chemistry, Faculty of Science, Masaryk University, Kotlarska 2, CZ-611 37 Brno, Czech Republic.
authors.forename: Libuse
authors.initials: L
authors.lastname: Trnkova
authors.affiliation: Department of Chemistry and Biochemistry Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, CZ-613 00 Brno, Czech Republic. kizek@sci.muni.cz.
authors.forename: Rene
authors.initials: R
authors.lastname: Kizek
keywords: Brdicka reaction; Proteomics; adsorptive transfer stripping technique; differential pulse voltammetry; human blood serum; metallothionein; thiols; tumour disease; zeptomole