Xu Chenjie, Xing Bengang, Rao Jianghong
Biophysics, Cancer Biology, and Molecular Imaging Programs, Department of Radiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Biochem Biophys Res Commun. 2006 Jun 9;344(3):931-5. doi: 10.1016/j.bbrc.2006.03.225. Epub 2006 Apr 19.
This communication describes a quantum dot probe that can be activated by a reporter enzyme, beta-lactamase. Our design is based on the principle of fluorescence resonance energy transfer (FRET). A biotinylated beta-lactamase substrate was labeled with a carbocyanine dye, Cy5, and immobilized on the surface of quantum dots through the binding of biotin to streptavidin pre-coated on the quantum dots. In assembling this nanoprobe, we have found that both the distance between substrates and the quantum dot surface, and the density of substrates are important for its function. The fluorescence emission from quantum dots can be efficiently quenched (up to 95%) by Cy5 due to FRET. Our final quantum dot probe, assembled with QD605 and 1:1 mixture of biotin and a Cy5-labeled lactam, can be activated by 32microg/mL of beta-lactamase with 4-fold increase in the fluorescence emission.
本通讯描述了一种可由报告酶β-内酰胺酶激活的量子点探针。我们的设计基于荧光共振能量转移(FRET)原理。一种生物素化的β-内酰胺酶底物用花青染料Cy5标记,并通过生物素与预涂覆在量子点上的链霉亲和素的结合固定在量子点表面。在组装这种纳米探针时,我们发现底物与量子点表面之间的距离以及底物的密度对其功能都很重要。由于FRET,量子点的荧光发射可被Cy5有效淬灭(高达95%)。我们最终的量子点探针由QD605与生物素和Cy5标记的内酰胺的1:1混合物组装而成,可被32μg/mL的β-内酰胺酶激活,荧光发射增加4倍。