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首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >Development of homogeneous binding assays based on fluorescence resonance energy transfer between quantum dots and Alexa Fluor fluorophores
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Development of homogeneous binding assays based on fluorescence resonance energy transfer between quantum dots and Alexa Fluor fluorophores

机译：基于量子点和Alexa Fluor荧光团之间荧光共振能量转移的均相结合测定方法的开发

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摘要

We studied the fluorescence resonance energy transfer (FRET) between quantum dots emitting at 565, 605, and 655 nm as energy donors and Alexa Fluor fluorophores with absorbance maxima at 594, 633, 647, and 680 nm as energy acceptors. As a first step, we prepared covalent conjugates between all three types of quantum dots and each of the Alexa, Fluor fluorophores that could act as an energy acceptor. All of these conjugates displayed efficient resonance energy transfer. Then we prepared covalent conjugates of these quantum dots with biotin, fluorescein, and cortisol and established that the binding of these conjugates to suitable Alexa Fluor-labeled antibodies and streptavidin (in the case of biotin) can be efficiently detected by measuring the resonance energy transfer in homogeneous solutions. Finally, based on these observations, competitive binding assays for these three small analytes were developed. The performance of these assays as a function of the degree of labeling of the quantum dots was evaluated. It was found that decreasing the degree of loading of the quantum dots leads to decreases of the limits of detection. The results show the great potential of this FRET system for the development of new homogeneous binding assays. (c) 2006 Elsevier Inc. All rights reserved.
机译：我们研究了在565、605和655 nm处作为能量供体发射的量子点与在594、633、647和680 nm处具有最大吸光度的Alexa Fluor荧光团之间的荧光共振能量转移（FRET）。第一步，我们准备了所有三种类型的量子点与可用作能量受体的Alexa Fluor荧光团之间的共价共轭物。所有这些共轭物均显示出有效的共振能量转移。然后，我们制备了这些量子点与生物素，荧光素和皮质醇的共价共轭物，并确定可以通过测量共振能量转移来有效检测这些共轭物与合适的Alexa Fluor标记抗体和链霉亲和素的结合（在生物素的情况下）在均匀溶液中。最后，基于这些观察结果，开发了针对这三种小分析物的竞争性结合测定。评估这些测定的性能随量子点标记程度的变化。发现降低量子点的负载度导致检测极限的降低。结果表明，该FRET系统具有开发新的均相结合测定的巨大潜力。 （c）2006 Elsevier Inc.保留所有权利。

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