Singh Bipin K, Hillier Andrew C
Department of Chemical Engineering, Iowa State University, Ames, Iowa 50011, USA.
Anal Chem. 2006 Mar 15;78(6):2009-18. doi: 10.1021/ac0519209.
A surface plasmon resonance sensor array based upon a grating substrate was developed for the detection of biomolecular interactions. The substrate consisted of a gold grating prepared by wet chemical treatment of a commercial recordable compact disk. A custom-built floating pin microspotter was constructed to deliver solutions containing omega-functionalized linear alkanethiols to the grating surface and produce an array of sensor elements with different exposed functional end groups. This array platform can be used to study biomolecular interactions in a label-free, sensitive, and high-throughput format. To illustrate the performance of this device, a test protein (bovine serum albumin) was exposed to sensor elements containing an array of functionalized alkanethiols possessing either activated carboxylic acid-, amine-, or hydroxyl-terminated regions. Local changes in plasmon resonance were monitored in a fixed-angle imaging configuration. Plasmon images clearly distinguish the degree of protein attachment at the various surfaces. The molecular binding events on the grating were also confirmed by ellipsometry. This grating-based SPR imaging platform represents a simple and robust method for performing label-free, high-sensitivity, and high-throughput detection of biomolecular interactions.
为检测生物分子相互作用,开发了一种基于光栅基底的表面等离子体共振传感器阵列。该基底由通过对商用可记录光盘进行湿化学处理制备的金光栅组成。构建了一个定制的浮动针微量点样器,用于将含有ω-官能化直链烷硫醇的溶液输送到光栅表面,并产生一系列具有不同暴露官能端基的传感器元件。该阵列平台可用于以无标记、灵敏且高通量的形式研究生物分子相互作用。为说明该装置的性能,将一种测试蛋白(牛血清白蛋白)暴露于含有一系列具有活化羧酸、胺或羟基末端区域的官能化烷硫醇的传感器元件。在固定角度成像配置下监测等离子体共振的局部变化。等离子体图像清楚地区分了蛋白质在不同表面的附着程度。通过椭偏仪也证实了光栅上的分子结合事件。这种基于光栅的表面等离子体共振成像平台代表了一种用于进行生物分子相互作用的无标记、高灵敏度和高通量检测的简单且稳健的方法。