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Selection of high affinity aptamer-ligand for dexamethasone and its electrochemical biosensor

机译:地塞米松高亲和力适体配体的选择及其电化学生物传感器

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摘要

A high specificity aptamer-ligand biorecognition and binding system to monitor of dexamethasone (DXN) was developed. The detection principle was based on a label-free electrochemical aptasensor. The selection of the aptamer was successfully performed by the systematic evolution of ligands through exponential enrichment technique (SELEX). From a random library of 1.08 × 1015 single-stranded DNA, an aptamer designated as DEX04 showed a highest affinity with a dissociation constant of 18.35 nM. It also showed a good conformational change when binding with DXN. In addition, the aptamer DEX04 did not show any cross-reactivity with other commonly used hormones. An impedimetric aptasensor for DXN was then developed by immobilizing DEX04 on a gold electrode. The binding upon to DXN was monitored by following the change in the charge transfer resistance (Rct) of the [Fe(CN)6]4−/3− redox couple. The aptasensor exhibited a linear range from 2.5 to 100 nM with a detection limit of 2.12 nM. When applied aptasensor to test in water samples, it showed good recovery percentages. The new DXN aptamer can be employed in other biosensing applications for food control and the diagnosis of some diseases in medicine as a cost-effective, sensitive and rapid detection method.
机译:开发了一种高特异性的适体-配体生物识别和结合系统来监测地塞米松(DXN)。检测原理基于无标记电化学适体传感器。通过指数富集技术(SELEX)通过配体的系统进化成功地进行了适体的选择。从1.08×10 15 单链DNA的随机文库中,命名为DEX04的适体显示出最高的亲和力,解离常数为18.35 nM。与DXN结合时,它还显示出良好的构象变化。另外,适体DEX04与其他常用激素没有显示出任何交叉反应。然后通过将DEX04固定在金电极上来开发用于DXN的阻抗适体传感器。通过跟踪[Fe(CN)6] 4- / 3-氧化还原对的电荷转移电阻(Rct)的变化来监控与DXN的结合。适体传感器的线性范围为2.5至100 nM,检出限为2.12 nM。当将aptasensor用于水样测试时,它显示出良好的回收率。新型DXN适体可作为具有成本效益,灵敏且快速的检测方法,用于食品控制和医学中某些疾病的诊断的其他生物传感应用中。

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