Signaling aptamer/protein binding by a molecular light switch complex
Signaling aptamer/protein binding by a molecular light switch complex
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DOI：
10.1021/ac049565u
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发表时间：
2004-09-01
影响因子：
7.4
通讯作者：
Bai, CL
中科院分区：
文献类型：
--
作者：
Jiang, YX;Fang, XH;Bai, CL
利用分子光开关配合物[Ru（phen）（2）-（dppz）]（2+），建立了一种新的适体/蛋白质结合信号的方法，用于基于适体的蛋白质检测。该方法利用了[Ru（phen）2（dppz）]2+在蛋白质/适体结合时嵌入适体的灵敏发光信号变化。首先测试针对免疫球蛋白E（IgE）的37-nt DNA适体作为模型系统。[Ru（phen）（2）（dppz）]（2+）/IgE适体的发光随IgE的增加而减弱。通过监测发光变化，我们能够检测适体和IgE之间的结合事件，用于在均质溶液以及血清中的IgE定量。该方法具有高度的选择性和灵敏度，检测限为100 pM的IgE。这种新方法非常简单，不需要将荧光团共价偶联到适体上。通过直接检测另外两种蛋白质，使用其DNA适体的癌蛋白血小板衍生生长因子-BB（PDGF-BB）和使用其RNA适体的α-凝血酶，证明了该方法的普遍性。这种新方法有望促进基于适体的生物传感器在生物化学和生物医学研究中的蛋白质测定的开发。
A novel method of signaling aptamer/protein binding for aptamer-based protein detection has been developed using a molecular light switch complex, [Ru(phen)(2)-(dppz)](2+). The method takes advantage of the sensitive luminescence signal change of [Ru(phen)2(dppz)]2+ intercalating to the aptamer upon protein/aptamer binding. A 37-nt DNA aptamer against immunoglobulin E (IgE) was first tested as a model system. The luminescence of the [Ru(phen)(2)(dppz)](2+)/IgE aptamer decreased with the increase of IgE. By monitoring the luminescence change, we were able to detect the binding events between the aptamer and IgE for IgE quantitation in homogeneous solutions as well as in serum. The assay was highly selective and sensitive with a detection limit of 100 pM for IgE. This new method is very simple and without the need for the covalent coupling of fluorophores to aptamers. The generalizability of the method was demonstrated by the direct detection of two other proteins, oncoprotein platelet derived growth factor-BB (PDGF-BB) using its DNA aptamer and a-thrombin using its RNA aptamer. This new approach is expected to promote the exploitation of aptamer-based biosensors for protein assays in biochemical and biomedical studies.