Murata Masaharu, Gouda Chifumi, 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 Oct;19(10):1355-7. doi: 10.2116/analsci.19.1355.
In vitro screening assays are useful techniques for the determination of receptor-mediated activities in environmental samples. In order to define whether environmental chemicals act as an agonist or antagonist to the human estrogen receptor (hER), we have constructed a biosensor based on ligand-inducible interactions between hER and relative proteins on a quartz crystal microbalance (QCM). The his-tagged proteins, which were expressed in E. coli by recombinant DNA technology, were immobilized on an Au-electrode with Ni(II)-mediated chemisorption using the histidine tag and thiol-modified iminodiacetic acid. The resonance-frequency change of the protein-modified electrode was caused by association or dissociation with the hER relative proteins on the surface in the presence of estrogen. These results suggest that this sensor is applicable as a large-scale screening tool for estrogenic compounds.
体外筛选测定法是测定环境样品中受体介导活性的有用技术。为了确定环境化学物质是人类雌激素受体（hER）的激动剂还是拮抗剂，我们基于hER与石英晶体微天平（QCM）上相关蛋白质之间的配体诱导相互作用构建了一种生物传感器。通过重组DNA技术在大肠杆菌中表达的带有组氨酸标签的蛋白质，利用组氨酸标签和硫醇修饰的亚氨基二乙酸，通过镍（II）介导的化学吸附固定在金电极上。在雌激素存在的情况下，蛋白质修饰电极的共振频率变化是由其与表面hER相关蛋白质的结合或解离引起的。这些结果表明，该传感器可作为雌激素化合物的大规模筛选工具。
