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基于太赫兹适体生物传感器的微囊藻毒素-LR 环藻毒素的高灵敏度小分子检测。

High-sensitivity small-molecule detection of microcystin-LR cyano-toxin using a terahertz-aptamer biosensor.

机构信息

Institut National de la Recherche Scientifique - Énergie Matériaux Télécommunications, Varennes, Québec J3X 1S2, Canada.

INRS Centre Armand-Frappier Santé Biotechnologie, 531 boulevards des Prairies, Laval, Québec, Canada.

出版信息

Analyst. 2021 Dec 6;146(24):7583-7592. doi: 10.1039/d1an01577j.

Abstract

We demonstrate the rapid and highly sensitive detection of a small molecule, microcystin-LR (MC-LR) toxin using an aptasensor based on a terahertz (THz) emission technique named the terahertz chemical microscope (TCM). The main component of the TCM is the sensing plate, which consists of a thin silicon layer deposited on a sapphire substrate, with a natural SiO layer formed on the top of the Si layer. The DNA aptamer is linked to the oxidized top surface of the silicon layer by a one-step reaction (click chemistry) between the DBCO-labeled aptamer and an azido group that binds to the surface. Using density functional theory (DFT) calculations, the number of active sites on the surface has been estimated to be 3.8 × 10 cm. Aptamer immobilization and MC-LR binding have been optimized by adjusting the aptamer concentration and the binding buffer composition. When MC-LR binds with the DNA aptamer, it causes a change in the chemical potential at the surface of the sensing plate, which leads to a change in the amplitude of the THz signal. Compared with other bio-sensing methods such as surface plasmon resonance (SPR), TCM is a rapid assay that can be completed in 15 min (10 min incubation and 5 min data acquisition). Moreover, our results show that the aptamer-based TCM can detect MC-LR with an excellent detection limit of 50 ng L, which is 20 times more sensitive compared with SPR measurements of MC-LR.

摘要

我们展示了一种基于太赫兹(THz)发射技术的适体传感器(称为太赫兹化学显微镜(TCM)),可快速、高灵敏度地检测小分子微囊藻毒素-LR(MC-LR)毒素。TCM 的主要组成部分是传感板,它由沉积在蓝宝石衬底上的薄硅层组成,在硅层的顶部形成天然的 SiO 层。DNA 适体通过 DBCO 标记的适体与结合到表面的叠氮基团之间的一步反应(点击化学)连接到氧化的硅层的顶表面。通过密度泛函理论(DFT)计算,估计表面的活性位点数量为 3.8×10^13 cm^-2。通过调整适体浓度和结合缓冲液组成优化了适体固定化和 MC-LR 结合。当 MC-LR 与 DNA 适体结合时,它会导致传感板表面的化学势发生变化,从而导致 THz 信号的幅度发生变化。与表面等离子体共振(SPR)等其他生物传感方法相比,TCM 是一种快速分析方法,可在 15 分钟内完成(10 分钟孵育和 5 分钟数据采集)。此外,我们的结果表明,基于适体的 TCM 可以检测 MC-LR,检测限为 50ng L,比 SPR 测量 MC-LR 的灵敏度高 20 倍。

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