†Key Laboratory of Coastal Environmental Processes and Ecological Remediation, Shandong Provincial Key Laboratory of Coastal Environmental Processes, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China.
§University of Chinese Academy of Sciences, Beijing 100049, China.
ACS Appl Mater Interfaces. 2015 May 6;7(17):9118-27. doi: 10.1021/acsami.5b00908. Epub 2015 Apr 23.
Phycocyanin with important physiological/environmental significance has attracted increasing attention; versatile molecularly imprinted polymers (MIPs) have been applied to diverse species, but protein imprinting is still quite difficult. Herein, using phycocyanin as template via a sol-gel process, we developed a novel fluorescent probe for specific recognition and sensitive detection of phycocyanin by quantum dots (QDs) based mesoporous structured imprinting microspheres (SiO2@QDs@ms-MIPs), obeying electron-transfer-induced fluorescence quenching mechanism. When phycocyanin was present, a Meisenheimer complex would be produced between phycocyanin and primary amino groups of QDs surface, and then the photoluminescent energy of QDs would be transferred to the complex, leading to the fluorescence quenching of QDs. As a result, the fluorescent intensity of the SiO2@QDs@ms-MIPs was significantly decreased within 8 min, and accordingly a favorable linearity within 0.02-0.8 μM and a high detectability of 5.9 nM were presented. Excellent recognition specificity for phycocyanin over its analogues was displayed, with a high imprinting factor of 4.72. Furthermore, the validated probe strategy was successfully applied to seawater and lake water sample analysis, and high recoveries in the range of 94.0-105.0% were attained at three spiking levels of phycocyanin, with precisions below 5.3%. The study provided promising perspectives to develop fluorescent probes for convenient, rapid recognition and sensitive detection of trace proteins from complex matrices, and further pushed forward protein imprinting research.
藻蓝蛋白具有重要的生理/环境意义,引起了越来越多的关注;多功能的分子印迹聚合物(MIPs)已被应用于多种物种,但蛋白质印迹仍然相当困难。在此,我们通过溶胶-凝胶过程以藻蓝蛋白为模板,开发了一种新型荧光探针,用于基于量子点(QDs)的介孔结构印迹微球(SiO2@QDs@ms-MIPs)对藻蓝蛋白进行特异性识别和灵敏检测,遵循电子转移诱导荧光猝灭机制。当存在藻蓝蛋白时,藻蓝蛋白和 QDs 表面的伯氨基之间会产生 Meisenheimer 复合物,然后 QDs 的光致发光能量会转移到复合物中,导致 QDs 的荧光猝灭。结果,SiO2@QDs@ms-MIPs 的荧光强度在 8 分钟内显著降低,因此在 0.02-0.8 μM 范围内呈现出良好的线性关系和 5.9 nM 的高检测灵敏度。该探针对藻蓝蛋白及其类似物表现出优异的识别特异性,印迹因子高达 4.72。此外,该验证后的探针策略已成功应用于海水和湖水样品分析,在藻蓝蛋白的三个加标水平下,回收率在 94.0-105.0%范围内,精度低于 5.3%。该研究为开发方便、快速识别复杂基质中痕量蛋白质的荧光探针提供了有前景的思路,并进一步推动了蛋白质印迹研究。