Dutta Pampa, Hill Kasey, Datskos Panos G, Sepaniak Michael J
Department of Chemistry, University of Tennessee, 420 Buehler Hall, Knoxville, Tennessee 37996-1600, USA.
Lab Chip. 2007 Sep;7(9):1184-91. doi: 10.1039/b704723a. Epub 2007 Jun 22.
A nanomechanical transducer is developed to detect and screen endocrine disrupting chemicals (EDCs) combining fluidic sample injection and delivery with bioreceptor protein functionalized microcantilevers (MCs). The adverse affects of EDCs on the endocrine system of humans, livestock, and wildlife provides strong motivation for advances in analytical detection and monitoring techniques. The combination of protein receptors, which include estrogen receptor alpha (ER-alpha) and estrogen receptor beta (ER-beta), as well as monoclonal antibodies (Ab), with MC systems employing modified nanostructured surfaces provides for excellent nanomechanical response sensitivity and the inherent selectivity of biospecific receptor-EDC interactions. The observed ranking of binding interaction of the tested EDCs with ER-beta is diethylstilbestrol (DES) > 17-beta-estradiol > 17-alpha-estradiol > 2-OH-estrone > bisphenol A > p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) with measurements exhibiting intra-day RSDs of about 3%. A comparison of responses of three EDCs, which include 17-beta-estradiol, 17-alpha-estradiol, and 2-OH-estrone, with ER-beta and ER-alpha illustrates which estrogen receptor subtype provides the greatest sensitivity. Antibodies specific to a particular EDC can also be used for analyte specific screening. Calibration plots for a MC functionalized with anti-17-beta-estradiol Ab show responses in the range of 1 x 10(-11) through 1 x 10(-7) M for 17-beta-estradiol with a linear portion extending over two orders of magnitude in concentration.
开发了一种纳米机械换能器,将流体样品注入和输送与生物受体蛋白功能化微悬臂梁(MC)相结合,用于检测和筛选内分泌干扰化学物质(EDC)。EDC对人类、牲畜和野生动物内分泌系统的不利影响为分析检测和监测技术的进步提供了强大动力。蛋白质受体(包括雌激素受体α(ER-α)和雌激素受体β(ER-β))以及单克隆抗体(Ab)与采用修饰纳米结构表面的MC系统相结合,提供了出色的纳米机械响应灵敏度和生物特异性受体-EDC相互作用的固有选择性。所测试的EDC与ER-β的结合相互作用观察排名为己烯雌酚(DES)>17-β-雌二醇>17-α-雌二醇>2-羟基雌酮>双酚A>对,对'-二氯二苯二氯乙烯(对,对'-DDE),测量显示日内相对标准偏差约为3%。比较三种EDC(包括17-β-雌二醇、17-α-雌二醇和2-羟基雌酮)与ER-β和ER-α的响应,说明了哪种雌激素受体亚型具有最高灵敏度。针对特定EDC的抗体也可用于分析物特异性筛选。用抗17-β-雌二醇Ab功能化的MC的校准曲线显示,对于17-β-雌二醇,响应范围为1×10^(-11)至1×10^(-7) M,线性部分在浓度上延伸两个数量级。