Aslan Kadir, Malyn Stuart N, Geddes Chris D
Institute of Fluorescence, Laboratory for Advanced Medical Plasmonics, Medical Biotechnology Center, University of Maryland Biotechnology Institute, Baltimore, MD, 21201, USA.
J Immunol Methods. 2007 May 31;323(1):55-64. doi: 10.1016/j.jim.2007.02.010. Epub 2007 Mar 28.
The applicability of a new technique, Microwave-Accelerated Surface Plasmon-Coupled Luminescence (MA-SPCL) for fast and sensitive bioassays in buffer, serum and whole blood using quantum dots as luminescence reporters is demonstrated. In this regard, a model bioassay based on the well-known interactions of biotin and streptavidin is used. Using MA-SPCL, the bioassay was kinetically completed within 1 min with the use of low power microwave heating as compared to the identical bioassay which took in excess of 30 min to reach >95% completion at room temperature, a 30-fold increase in assay kinetics. The luminescence emission from the quantum dots was coupled to surface plasmons of the gold film, enabling the detection of the luminescence emission in a highly directional fashion as compared to the normal isotropic emission, for enhanced sensitivity and detection. The combined effect of microwaves for faster assay kinetics, with surface plasmon-coupled luminescence for sensitive luminescence measurements, has also made possible the demonstration of the use of the MA-SPCL technique for assays run in complex media, such as human serum and whole blood, where the same assay could not be performed at room temperature due to the coagulation of blood. In the MA-SPCL assay run in serum and whole blood, the luminescence intensity from 33 nM quantum dots was 75% and 20% that of the luminescence intensity from the assay run in buffer, with a signal to noise ratio of 12.5 and 3, respectively.
本文展示了一种新技术——微波加速表面等离子体耦合发光(MA-SPCL)在缓冲液、血清和全血中使用量子点作为发光报告分子进行快速灵敏生物测定的适用性。在此方面,使用了基于生物素和链霉亲和素著名相互作用的模型生物测定。与在室温下超过30分钟才能达到>95%完成度的相同生物测定相比,使用MA-SPCL,通过低功率微波加热,该生物测定在1分钟内即可动力学完成,测定动力学提高了30倍。量子点的发光发射与金膜的表面等离子体耦合,与正常的各向同性发射相比,能够以高度定向的方式检测发光发射,从而提高灵敏度和检测能力。微波对更快测定动力学的作用与表面等离子体耦合发光对灵敏发光测量的作用相结合,也使得MA-SPCL技术可用于在复杂介质(如人血清和全血)中进行的测定得以展示,在室温下由于血液凝固无法进行相同的测定。在血清和全血中进行的MA-SPCL测定中,33 nM量子点的发光强度分别为在缓冲液中进行测定时发光强度的75%和20%,信噪比分别为12.5和3。