Department of Chemistry and the Advanced Materials Research Institute, University of New Orleans, New Orleans, Louisiana 70148, USA.
J Am Chem Soc. 2010 Feb 10;132(5):1460-1. doi: 10.1021/ja908784b.
This paper describes the development of new fluorescence resonance energy transfer (FRET)-based quantum dot probes for proteolytic activity. The CdSe/ZnS quantum dots are incorporated into a thin polymeric film, which is prepared by layer-by-layer deposition of alternately charged polyelectrolytes. The quantum dots, which serve as fluorescent donors, are separated from rhodamine acceptor molecules, which are covalently attached to the film surface by a varying number of polyelectrolyte layers. When excited with visible light, the emission color of the polyelectrolyte multilayer film appears orange due to FRET between the quantum dots and molecular acceptors. The emission color changes to green when the rhodamine molecules are removed from the surface by enzymatic cleavage. The new probe design enables the use of quantum dots in bioassays, in this study for real-time monitoring of trypsin activity, while alleviating concerns about their potential toxicity. Application of these quantum dot FRET-based probes in microfluidic channels enables bioanalysis of volume-limited samples and single-cell studies in an in vivo-like environment.
本文描述了用于蛋白酶活性的新型荧光共振能量转移（FRET）量子点探针的研制。CdSe/ZnS 量子点被掺入到由交替带电荷的聚合物层层沉积而成的薄膜中。量子点作为荧光供体，与通过不同数量的聚合物层共价连接到薄膜表面的罗丹明受体分子分离。当用可见光激发时，由于量子点和分子受体之间的 FRET，聚合物多层膜的发射颜色呈现橙色。当通过酶切从表面除去罗丹明分子时，发射颜色变为绿色。这种新的探针设计使得可以在生物测定中使用量子点，在本研究中用于实时监测胰蛋白酶活性，同时减轻了对其潜在毒性的担忧。这些基于量子点 FRET 的探针在微流道中的应用可以对体积有限的样品进行生物分析，并在类似于体内的环境中进行单细胞研究。
