适体
凝血酶
分析物
化学
荧光
检出限
动力学
组合化学
分析化学(期刊)
色谱法
分子生物学
血小板
物理
量子力学
免疫学
生物
作者
Xinling Zeng,Qing Zhou,Liyan Wang,Xiaoxian Zhu,Kuiyan Cui,Xinsheng Peng,Terry W. J. Steele,Huizhi Chen,Hui Xu,Yubin Zhou
出处
期刊:Materials
[MDPI AG]
日期:2021-11-16
卷期号:14 (22): 6927-6927
被引量:5
摘要
It is important to detect thrombin due to its physiological and pathological roles, where rapid and simple analytical approaches are needed. In this study, an aptasensor based on fluorescence attenuation kinetics for the detection of thrombin is presented, which incorporates the features of stilbene and aptamer. We designed and synthesized an aptasensor by one-step coupling of stilbene compound and aptamer, which employed the adaptive binding of the aptamer with thrombin to cause a change in stilbene fluorescence attenuation kinetics. The sensor realized detection of thrombin by monitoring the variation in apparent fluorescence attenuation rate constant (kapp), which could be further used for probing of enzyme-aptamer binding. In comprehensive studies, the developed aptasensor presented satisfactory performance on repeatability, specificity, and regeneration capacity, which realized rapid sensing (10 s) with a limit of detection (LOD) of 0.205 μM. The strategy was successful across seven variants of thrombin aptasensors, with tunable kapp depending on the SITS (4-Acetamido-4'-isothiocyanato-2,2'-stilbenedisulfonic acid disodium salt hydrate) grafting site. Analyte detection mode was demonstrated in diluted serum, requiring no separation or washing steps. The new sensing mode for thrombin detection paves a way for high-throughput kinetic-based sensors for exploiting aptamers targeted at clinically relevant proteins.
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