Adam Vojtech, Baloun Jiri, Fabrik Ivo, Trnkova Libuse, Kizek Rene
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.
Sensors (Basel). 2008 Apr 1;8(4):2293-2305. doi: 10.3390/s8042293.
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.
纳升体积中zeptomole水平金属硫蛋白的电化学检测
我们报道了吸附转移溶出技术(AdTS)与差分脉冲伏安法Brdicka反应相结合用于测定硫醇蛋白的改进方法。当前技术在分析极低样品体积(纳升)时无法产生可重复的结果。通过改进当前的转移技术,包括对吸附分析物进行冷却步骤,这一障碍在技术上可以克服。我们将该技术用于测定一种有前景的肿瘤疾病标志物蛋白——金属硫蛋白(MT)。MT的检测限(3S/N)评估为每500纳升500zeptomoles(1pM),定量限(10S/N)为每500纳升1500zeptomoles(3pM)。此外,改进后的AdTS技术被用于分析乳腺癌患者的血清样本。根据所得结果可以得出结论,改进后的技术可用于在极低样品体积中检测硫醇蛋白,并且还能防止在洗涤和转移步骤中出现干扰。