Electro-Optic Engineering Unit and Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Environmental Engineering Unit, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Biosensors (Basel). 2013 Mar 19;3(1):157-170. doi: 10.3390/bios3010157. eCollection 2013 Mar.
An immobilization protocol was developed to attach receptors on smooth silver thin films. Dense and packed 11-mercaptoundecanoic acid (11-MUA) was used to avoid uncontrolled sulfidization and harmful oxidation of silver nanolayers. N,N'-dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) were added to make the silver surfaces reactive. A comparative study was carried out with different immersion times of silver samples in 11-MUA solutions with different concentrations to find the optimum conditions for immobilization. The signals, during each step of the protocol, were analyzed with a refractometer based on the surface plasmon resonance (SPR) effect and luminescence techniques. Molecular interactions at the surfaces between the probe and target at the surface nanolayer shift the SPR signal, thus indicating the presence of the substance. To demonstrate specific biosensing, rabbit anti-estrone polyclonal immunoglobulin G (IgG) antibody was immobilized through a linker on 47 nm silver layer deposited on SF11 glass. At the final stage, the representative endocrine disruptor-estrone-was attached and detected in deionized water with a diverging beam SPR imaging sensor.
我们开发了一种固定化方案，用于将受体固定在光滑的银薄膜上。使用密集且包装的 11-巯基十一烷酸（11-MUA）可避免银纳米层不受控制的硫化和有害氧化。添加 N,N'-二环己基碳二亚胺（DCC）和 N-羟基琥珀酰亚胺（NHS）可使银表面具有反应性。通过在不同浓度的 11-MUA 溶液中浸泡不同时间的银样品进行了比较研究，以找到最佳固定化条件。该方案的每个步骤的信号都通过基于表面等离子体共振（SPR）效应和发光技术的折射计进行分析。探针和目标在表面纳米层之间的分子相互作用会改变 SPR 信号，从而表明物质的存在。为了证明特定的生物传感，通过连接子将兔抗雌酮多克隆免疫球蛋白 G（IgG）抗体固定在沉积在 SF11 玻璃上的 47nm 银层上。在最后阶段，将代表性的内分泌干扰物-雌酮-连接并在去离子水中用发散光束 SPR 成像传感器进行检测。
