CAS Key Laboratory of Coastal Environmental Processes and Ecological Remediation, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China.
Analyst. 2018 Jul 23;143(15):3570-3578. doi: 10.1039/c8an00811f.
Versatile molecular imprinted fluorescence sensors have been prepared for various species, but the imprinting based fluorescence detection of target proteins upon different external stimulation is rarely reported. Herein, a novel phycocyanin-imprinted ratiometric fluorescence nanosensor was developed for the temperature regulated sensing and detection of a phycocyanin target based on fluorescence resonance energy transfer (FRET). The nanosensor was fabricated via simple facile copolymerization, with amino/carboxyl modified quantum dots (QDs) as a fluorescent support, N-isopropylacrylamide as a thermo-responsive functional monomer, N,N-methylenebisacrylamide as a cross-linker and phycocyanin as a template. Under temperature control at 20 °C and 45 °C, the fluorescence intensities of the QDs and phycocyanin were regularly decreased and enhanced, respectively, to a different extent as the concentration of phycocyanin increased, and thereby the ratio of the two fluorescence peak emission intensities of QDs and phycocyanin was used to determine the concentration of phycocyanin. Good linearity was obtained within the range of 0-1.8 μM (r = 0.9900) with a low detection limit of 3.2 nM, and excellent recognition selectivity towards the phycocyanin target was achieved over other proteins. Moreover, satisfactory recoveries of 92.0-106.8% were obtained in spiked seawater samples. This study provides a facile, fast and intelligent way to conduct identification analysis of trace proteins in complex water matrices, and can push forward protein imprinting and stimuli-responsive imprinting related research.
已经制备了多功能分子印迹荧光传感器，用于各种物种，但很少有报道基于印迹的荧光检测不同外部刺激下的靶蛋白。本文报道了一种新型的藻蓝蛋白印迹比率荧光纳米传感器，用于基于荧光共振能量转移（FRET）的温度调控传感和藻蓝蛋白靶标检测。该纳米传感器通过简单的共聚反应制备，以氨基/羧基修饰的量子点（QDs）为荧光支撑体，N-异丙基丙烯酰胺为热敏功能单体，N，N-亚甲基双丙烯酰胺为交联剂，藻蓝蛋白为模板。在 20°C 和 45°C 的温度控制下，随着藻蓝蛋白浓度的增加，QDs 和藻蓝蛋白的荧光强度分别以不同的程度有规律地降低和增强，从而利用两者的荧光峰发射强度比来测定藻蓝蛋白的浓度。在 0-1.8 μM 范围内得到了良好的线性关系（r = 0.9900），检测限低至 3.2 nM，对其他蛋白质具有优异的识别选择性。此外，在加标海水中样品的回收率为 92.0-106.8%。本研究为在复杂水基质中进行痕量蛋白质的识别分析提供了一种简便、快速和智能的方法，并能推动蛋白质印迹和刺激响应印迹相关研究。
