Murata Masaharu, Yano Kentaro, Kuroki Shinichiro, Suzutani Tatsuo, Katayama Yoshiki
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Fukuoka 812-8581, Japan.
Anal Sci. 2003 Dec;19(12):1569-73. doi: 10.2116/analsci.19.1569.
The recombinant human thyroid hormone receptor (TR) was expressed as an in-frame fusion with ten consecutive histidine residues using a bacterial system; then the receptor was immobilized on an Au-electrode with Ni(II)-mediated chemisorption using the histidine tag and thiol-modified nitrilotriacetic acid. The receptor-modified electrode could rapidly detect ligand binding to hTR without any separation procedures, and showed a good response in a concentration-dependent manner. The sensitivity of this biosensor based on ligand-receptor interactions was comparable to those of conventional competitive ligand binding assays using radio-labeled ligands. Our results strongly suggest that our new biosensor can be applied to the identification of new ligands for hTR.
使用细菌系统将重组人甲状腺激素受体（TR）表达为与十个连续组氨酸残基的读框内融合体；然后利用组氨酸标签和硫醇修饰的次氮基三乙酸，通过镍（II）介导的化学吸附将该受体固定在金电极上。该受体修饰电极无需任何分离程序就能快速检测配体与hTR的结合，并且呈浓度依赖性地表现出良好的响应。这种基于配体-受体相互作用的生物传感器的灵敏度与使用放射性标记配体的传统竞争性配体结合测定法相当。我们的结果有力地表明，我们的新型生物传感器可应用于鉴定hTR的新配体。
