Copper-Mediated Amplification Allows Readout of Immunoassays by the Naked Eye
Copper-Mediated Amplification Allows Readout of Immunoassays by the Naked Eye
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铜介导的扩增允许肉眼读出免疫测定结果
DOI：
10.1002/anie.201006025
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发表时间：
2011-01-01
影响因子：
16.6
通讯作者：
Jiang, Xingyu
中科院分区：
文献类型：
--
作者：
Qu, Weisi;Liu, Yingyi;Jiang, Xingyu
本文介绍了一种新的标记抗体的方法，这种方法不需要使用先进的设备，就可用于免疫分析的比色检测。色度检测在许多应用中是方便和有效的，因为读出仅需要人眼。金纳米颗粒（Au NP）可用于比色测定，因为具有低浓度Au NP的溶液的聚集显示出清晰的颜色变化。溶液中Au NPs的聚集产生了许多测定，[1]包括离子，[2，3]小分子，[4] DNA，[5，6]蛋白质，[7]和癌细胞。[8，9] CuI催化的拴系反应的进展，例如室温下水溶液中的“点击化学”[10，11]，为基于Au NP的生化分析提供了新型化学。我们以前的工作表明，基于CuI的点击化学允许化学分析的高灵敏度和选择性[2]。使用铜作为催化剂（反应需要少量的铜）的事实确保了灵敏度，并且反应与大多数已知的化学反应正交的事实确保了选择性。在CuII与抗坏血酸钠作为还原剂的存在下，Au纳米粒子，具有叠氮化物和炔封端的基团进行聚集作为结果的CuI催化点击化学。在环境温度下，这个过程可以通过肉眼监测。即使在存在高浓度的其他阳离子和干扰分子的混合物的情况下，该方法也具有高度特异性。我们推断，如果这种检测方法可以扩展到检测CuII以外的物种，它通常可能适用于许多不同类型的高灵敏度和选择性检测。我们现在报告了一种基于检测从一氧化二铜纳米颗粒（CuO NP）-标记的抗体释放的CuII作为二抗的比色免疫测定法，代替传统上用于免疫测定的荧光染料或酶标记的二抗（方案1）。
A new method of labeling antibodies for the colorimetric detection of immunoassays without using advanced equipment is described. Colorimetric detections are convenient and effective in many applications because the readout requires only human eyes. Gold nanoparticles (Au NPs) are useful in colorimetric assays because the aggregation of solutions with low concentrations of Au NPs displays a clear color change. The aggregation of Au NPs in solution has yielded many assays,[1] including those for ions,[2, 3] small molecules,[4] DNA,[5, 6] proteins,[7] and cancerous cells.[8, 9] Advances in CuI-catalyzed tethering reactions, such as “click chemistry”[10, 11] in aqueous solutions at room temperature, have allowed for new types of chemistry for Au NP-based biochemical analysis.Our previous work shows that CuI-based click chemistry allows high sensitivity and selectivity [2] for chemical analysis. The fact that copper is used as a catalyst (a small amount of it is required for the reaction) ensures the sensitivity, and the fact that the reaction is orthogonal to most known chemical reactions ensures the selectivity. In the presence of CuII with sodium ascorbate as the reductant, Au NPs that have azideand alkyne-terminated groups undergo aggregation as the result of CuI-catalyzed click chemistry. At ambient temperature, this process can be monitored by the naked eye. This method is highly specific even in the presence of high concentrations of mixtures of other cations and interfering molecules. We reasoned that if such assays can be extended to detect species other than CuII, it might be generally useful for many different types of highly sensitive and selective assays. We now report a colorimetric immunoassay based on detecting CuII released from copper monoxide nanoparticle (CuO NP)-labeled antibodies as the secondary antibody, in place of the fluorescent dye-or enzyme-labeled secondary antibodies traditionally used in immunoassays (Scheme 1).