Ikebukuro Kazunori, Kiyohara Chiharu, Sode Koji
Department of Life Science and Biotechnology, Tokyo University of Agriculture and Technology, Nakamachi 2 24 16, Koganei, Tokyo 184 8588, Japan.
Biosens Bioelectron. 2005 Apr 15;20(10):2168-72. doi: 10.1016/j.bios.2004.09.002.
Novel electrochemical detection system for protein in sandwich manner using the aptamers was developed. Two different aptamers, which recognize different positions of thrombin, were chosen to construct sandwich type sensing system for protein, and one was immobilized onto the gold electrode for capturing thrombin onto the electrode and the other was used for detection. To obtain the signal, the aptamer for detection was labeled with pyrroquinoline quinone glucose dehydrogenase ((PQQ)GDH), and the electrical current, generated from glucose addition after the formation of the complex of thrombin, gold immobilized aptamer and the (PQQ)GDH labeled aptamer on the electrode, was measured. The increase of the electric current generated by (PQQ)GDH was observed in dependent manner of the concentration of thrombin added, and more than 10nM thrombin was detected selectively. The batch type protein sensing system was constructed using the two different aptamers sandwiching thrombin and it showed linear response to the increase of the thrombin concentration in the range of 40-100 nM.
开发了一种新颖的基于适体的夹心式蛋白质电化学检测系统。选择两种识别凝血酶不同位置的不同适体,构建蛋白质夹心型传感系统,其中一种固定在金电极上用于将凝血酶捕获到电极上,另一种用于检测。为了获得信号,用于检测的适体用吡咯喹啉醌葡萄糖脱氢酶((PQQ)GDH)标记,在凝血酶、固定有金的适体和电极上(PQQ)GDH标记的适体形成复合物后加入葡萄糖所产生的电流被测量。观察到(PQQ)GDH产生的电流增加与添加的凝血酶浓度呈依赖关系,并且能够选择性地检测到超过10 nM的凝血酶。使用两种不同的适体夹心法构建了批式蛋白质传感系统,它在40 - 100 nM范围内对凝血酶浓度的增加呈现线性响应。