Wittliff James L, Andres Sarah A, Kruer Traci L, Kerr D Alan, Smolenkova Irina A, Erb Judith L
Department of Biochemistry & Molecular Biology, Institute for Molecular Diversity & Drug Design, University of Louisville, Louisville, KY, USA.
Adv Exp Med Biol. 2008;614:315-22. doi: 10.1007/978-0-387-74911-2_35.
A novel evanescent-based biosensor (Endotect, ThreeFold Sensors, Inc.) was developed with laser-based fiber optics using fluorescent dye-labeled recombinant human estrogen receptor-alpha (rhERalpha) and hERbeta as probes. A three-tiered approach evaluating various steps in the formation of the estrogen-receptor complex and its subsequent activity was developed for instrument calibration to detect estrogen mimics in biological samples, water and soil. Using this approach, binding affinities and activities of certain known estrogen mimics were determined for their use as calibrator molecules. Results indicated rhERalpha and rhERbeta may be employed as probes to distinguish estrogen mimics with a broad range of affinities. In addition, application of the biosensor for detecting DNA-binding proteins in human tissue extracts was demonstrated. The later studies suggest the biosensor may be used as a clinical laboratory tool for assessing tumor marker proteins.
一种新型的基于倏逝波的生物传感器(Endotect,ThreeFold Sensors公司)通过基于激光的光纤技术开发而成,使用荧光染料标记的重组人雌激素受体α(rhERα)和hERβ作为探针。开发了一种三层方法,用于评估雌激素受体复合物形成过程中的各个步骤及其后续活性,以进行仪器校准,从而检测生物样品、水和土壤中的雌激素模拟物。使用这种方法,确定了某些已知雌激素模拟物的结合亲和力和活性,以便用作校准分子。结果表明,rhERα和rhERβ可用作探针,以区分具有广泛亲和力的雌激素模拟物。此外,还展示了该生物传感器在检测人体组织提取物中DNA结合蛋白方面的应用。后续研究表明,该生物传感器可用作评估肿瘤标志物蛋白的临床实验室工具。