Bendikov Tatyana A, Rabinkov Aharon, Karakouz Tanya, Vaskevich Alexander, Rubinstein Israel
Department of Materials and Interfaces, Weizmann Institute of Science, Rehovot 76100, Israel.
Anal Chem. 2008 Oct 1;80(19):7487-98. doi: 10.1021/ac8013466. Epub 2008 Aug 29.
Discontinuous, island-type gold films (typically < or = 10 nm nominal thickness) prepared by evaporation of the metal on transparent substrates show a localized surface plasmon resonance (LSPR) extinction in the visible-to-NIR range and can be used as optical transducers for monitoring local refractive index change. Such transducers, operated in the transmission configuration, provide an effective scheme for label-free biological sensing using basic spectrophotometric equipment. Optimization of the sensitivity of LPSR transducers requires consideration of the distance between the metal island surface and the bound analyte, strongly affecting the optical response due to the fast decay of the evanescent field of localized plasmons. In the present work Au island based LSPR transducers were used to monitor antibody-antigen interactions, demonstrating the effect of the biorecognition interface thickness. Evaporated Au island films derivatized with IgG or hCG antigens were used as biological recognition elements for selective sensing of antibody binding, distinguishing between specific and nonspecific interactions. The LSPR results are supported by XPS and ellipsometry data as well as by AFM and HRSEM imaging, the latter providing actual visualization of the two protein binding steps. Increase of the recognition interface thickness leads to a concomitant decrease in the extinction and wavelength sensitivity, generally conforming to a model of an exponentially decaying surface plasmon (SP) evanescent field.
通过在透明基板上蒸发金属制备的不连续岛状金膜(通常标称厚度≤10 nm)在可见光至近红外范围内呈现局域表面等离子体共振(LSPR)消光,并且可用作监测局部折射率变化的光学传感器。这种以透射配置运行的传感器,利用基本的分光光度计设备,为无标记生物传感提供了一种有效的方案。优化LPSR传感器的灵敏度需要考虑金属岛表面与结合分析物之间的距离,由于局域等离子体的倏逝场快速衰减,该距离会强烈影响光学响应。在本工作中,基于金岛的LSPR传感器用于监测抗体 - 抗原相互作用,证明了生物识别界面厚度的影响。用IgG或hCG抗原衍生化的蒸发金岛膜用作生物识别元件,用于选择性检测抗体结合,区分特异性和非特异性相互作用。LSPR结果得到XPS和椭偏测量数据以及AFM和HRSEM成像的支持,后者提供了两个蛋白质结合步骤的实际可视化。识别界面厚度的增加导致消光和波长灵敏度随之降低,这通常符合表面等离子体(SP)倏逝场指数衰减的模型。