Department of Materials, Imperial College London, London, UK.
Nat Protoc. 2013 Sep;8(9):1759-64. doi: 10.1038/nprot.2013.085. Epub 2013 Aug 22.
This protocol describes a signal-generation mechanism for the naked-eye detection of analytes at low concentrations with ELISA. The key step is to generate solutions of desired tonality by growing gold nanoparticles with a particular state of aggregation. This is accomplished by linking the growth of gold nanoparticles with the biocatalytic cycle of the enzyme label. The protocol adapts a conventional ELISA procedure with catalase-labeled antibodies. The enzyme consumes hydrogen peroxide, and then gold (III) ions are added to generate gold nanoparticles. The concentration of hydrogen peroxide dictates the state of aggregation of gold nanoparticles. This allows for the naked-eye detection of analytes by observing the generation of blue- or red-colored gold nanoparticle solutions. When coupled with conventional ELISA, this signal-generation procedure allows for the naked-eye detection of analytes within 1 h.
本方案描述了一种利用 ELISA 进行低浓度分析物的肉眼检测的信号产生机制。关键步骤是通过生长具有特定聚集状态的金纳米粒子来生成所需色调的溶液。这是通过将金纳米粒子的生长与酶标记的生物催化循环相联系来实现的。该方案采用了一种带有过氧化物酶标记抗体的常规 ELISA 程序。该酶消耗过氧化氢，然后加入金（III）离子以生成金纳米粒子。过氧化氢的浓度决定了金纳米粒子的聚集状态。这使得可以通过观察蓝色或红色金纳米粒子溶液的生成来进行分析物的肉眼检测。当与常规 ELISA 结合使用时，这种信号产生程序可以在 1 小时内实现分析物的肉眼检测。
