Quantum dot/bioluminescence resonance energy transfer based highly sensitive detection of proteases
Quantum dot/bioluminescence resonance energy transfer based highly sensitive detection of proteases
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DOI：
10.1002/anie.200700280
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发表时间：
2007-01-01
影响因子：
16.6
通讯作者：
Rao, Jianghong
中科院分区：
文献类型：
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作者：
Yao, Hequan;Zhang, Yan;Rao, Jianghong
半导体量子点（QDs）是一种具有高量子产率、高摩尔消光系数、尺寸相关可调谐发射和高光稳定性的明亮荧光发射体。[1-6]这些吸引人的荧光特性促使人们对开发基于量子点的传感器用于生物检测和成像产生了广泛的兴趣。[7-13]一种常用的开发量子点纳米传感器的策略是基于荧光共振能量转移（FRET），量子点作为FRET供体。有许多基于fret的QD生物传感器的例子，包括用于检测麦芽糖、pH、2,4,6 -三硝基甲苯、凝血酶和酶活性的自组装纳米复合物。[7-13]在这些基于FRET的量子点纳米传感器中，FRET受体的多个拷贝通常存在于一个量子点上，这可能导致自猝灭并导致FRET受体的低发射我们最近证明了量子点也可以作为发光蛋白（例如，生物发光蛋白Renilla荧光素酶）在生物发光共振能量转移（BRET）中的能量受体。[15,16]当量子点共轭物暴露于荧光素酶底物时，底物氧化释放的能量通过BRET传递给量子点，从而从量子点产生发光。QD BRET已被证明对体内成像和检测具有很高的灵敏度。在此，我们报道了基于abret的QD生物传感器的设计，用于检测基质金属蛋白酶（MMPs）的活性。MMPs是一个依赖锌的分泌内肽酶家族，对细胞外基质的调节降解和加工至关重要，并且在几乎所有类型的人类癌症中都上调。[17,18] MMPs在促进癌症进展中的重要作用使其成为药物开发和体内肿瘤检测的重要靶点。基于荧光和磁共振的方法已被用于检测MMPs的活性。[19-23]在这里，我们描述了一种基于bret的QD纳米传感器，用于高灵敏度检测明胶酶MMP-2的活性。
Semiconductor quantum dots (QDs) are bright fluorescence emitters with high quantum yields, high molar extinction coefficients, size-dependent tunable emission, and high photostability.[1–6] These attractive fluorescence properties prompt a wide interest in developing QD-based sensors for biological detection and imaging.[7–13] One often-used strategy towards the development of QD nanosensors is based on fluorescence resonance energy transfer (FRET) with the QDs as the FRET donor. There are numerous examples of FRET-based QD biosensors that include self-assembled nanocomplexes for detecting maltose, pH, 2, 4, 6-trinitrotoluene, thrombin, and enzyme activity.[7–13] In these FRET-based QD nanosensors, multiple copies of the FRET acceptor were often present on one QD, which may result in self-quenching and lead to low emission from the FRET acceptor.[14] We have recently demonstrated that QDs can also serve as an energy acceptor for a light-emitting protein (for example, the bioluminescent protein Renilla luciferase) in bioluminescence resonance energy transfer (BRET).[15, 16] When the QD conjugates are exposed to the luciferase substrate, the energy released in the oxidation of the substrate is transferred to the QDs through BRET, thus generating light emission from the QDs. The QD BRET has been shown to have high sensitivity for in vivo imaging and detection. Herein, we report the design of aBRET-based QD biosensor for detection of the activity of matrix metalloproteinases (MMPs). MMPs are a family of zinc-dependent secreted endopeptidases that are crucial for the regulated degradation and processing of extracellular matrices, and are upregulated in almost every type of human cancer.[17, 18] The significant role of MMPs in promoting cancer progression makes them important targets for drug development and in vivo tumor detection. Fluorescence-and magnetic-resonance-based approaches have been used to detect the activity of MMPs.[19–23] Here, we describe a BRET-based QD nanosensor for detecting the activity of gelatinase MMP-2 with high sensitivity.