College of Chemistry, Chemical Engineering and Materials Science, Soochow University , Suzhou 215123, China.
ACS Appl Mater Interfaces. 2013 Oct 9;5(19):9798-802. doi: 10.1021/am4029735. Epub 2013 Sep 20.
We describe here a simple fluorometric assay for the highly sensitive detection of caspase-3 activities on the basis of the inner-filter effect of gold nanoparticles (AuNPs) on CdTe quantum dots (QDs). The method takes advantage of the high molar absorptivity of the plasmon band of gold nanoparticles as well as the large absorption band shift from 520 to 680 nm upon nanoparticle aggregation. When labeled with a peptide possessing the caspase-3 cleavage sequence (DEVD), the monodispersed Au-Ps (peptide-modified AuNPs) exhibited a tendency to aggregate when exposed to caspase-3, which induced the absorption band transition from 520 to 680 nm and turned on the fluorescence of the CdTe QDs for caspase-3 sensing. Under optimum conditions, a high sensitivity towards caspase-3 was achieved with a detection limit as low as 18 pM, which was much lower than the corresponding assays based on absorbance or other approaches. Overall, we demonstrated a facile and sensitive approach for caspase-3 detection, and we expected that this method could be potentially generalized to design more fluorescent assays for sensing other bioactive entities.
我们在这里描述了一种基于金纳米粒子(AuNPs)对内滤效应CdTe 量子点(QDs)的高度灵敏的 caspase-3 活性的荧光检测方法。该方法利用了金纳米粒子等离子体带的高摩尔吸光率以及纳米颗粒聚集时从 520nm 到 680nm 的大吸收带移动。当用具有半胱天冬酶-3 切割序列(DEVD)的肽标记时,暴露于半胱天冬酶-3 时,单分散的 Au-Ps(肽修饰的 AuNPs)表现出聚集的趋势,这诱导了从 520nm 到 680nm 的吸收带跃迁,并开启了 CdTe QDs 的荧光用于半胱天冬酶-3 的感测。在最佳条件下,对 caspase-3 实现了高灵敏度,检测限低至 18 pM,远低于基于吸光度或其他方法的相应测定法。总的来说,我们展示了一种用于 caspase-3 检测的简单而灵敏的方法,我们期望该方法可以被推广到设计用于检测其他生物活性实体的更多荧光测定法。