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简易诊断:用于多巴胺捕获及通过双色比色和荧光系统进行准确检测的适配芯片

Diagnosis by simplicity: an aptachip for dopamine capture and accurate detection with a dual colorimetric and fluorometric system.

作者信息

Lin Tzu-Yang, Wei Kuo-Chen, Ju Shin-Pon, Huang Chiung-Yin, Yang Hung-Wei

机构信息

Institute of Medical Science and Technology, National Sun Yat-sen University, 70 Lienhai Rd., Kaohsiung, 80424, Taiwan.

出版信息

J Mater Chem B. 2018 May 28;6(20):3387-3394. doi: 10.1039/c8tb00913a. Epub 2018 May 16.

Abstract

In this study, we aim to rapidly fabricate an aptachip with a dual colorimetric and fluorometric sensing strategy for easy dopamine (DA) detection with high sensitivity and selectivity. To construct an aptachip with high DA capture efficiency, molecular dynamics (MD) simulations were utilitized to predict the most stable configuration of the DA-binding aptamer (DBA) for DA recognition. The DA in the specimen would be specifically captured on the DBA-aptachip, then released from the DBA in alkaline solution to form DA-quinone (DAQ), thus leading to a color change (from colorless to brown) and inducing a dramatic decrease in the fluorescence intensity as a result of the photoinduced electron transfer (PET) for bovine serum albumin (BSA)-stabilized Au nanoclusters (BSA-Au NCs). The detection limit of DA is as low as 0.1 μg mL for the colorimetric system and 0.5 ng mL for the fluorometric system. In addition, this biosensing of DA is easy to implement for visual detection owing to the DA oxidation and fluorescence quenching by BSA-AuNCs in the presence of the alkaline solution. Both the colorimetric and fluorometric systems showed excellent selectivity toward DA over interfering substances. Furthermore, we demonstrated the application of the present approach to analysis of artificial cerebrospinal fluid (ACSF) and serum samples, suggesting that this system holds promise for diagnostics.

摘要

在本研究中,我们旨在快速制备一种具有双色比色和荧光传感策略的适配体芯片,以便于高灵敏度和高选择性地检测多巴胺(DA)。为构建具有高DA捕获效率的适配体芯片,利用分子动力学(MD)模拟来预测用于DA识别的DA结合适配体(DBA)的最稳定构型。样本中的DA将被特异性捕获在DBA-适配体芯片上,然后在碱性溶液中从DBA释放形成多巴胺醌(DAQ),从而导致颜色变化(从无色变为棕色),并由于牛血清白蛋白(BSA)稳定的金纳米簇(BSA-Au NCs)的光致电子转移(PET)而导致荧光强度急剧下降。比色系统中DA的检测限低至0.1μg/mL,荧光系统中为0.5ng/mL。此外,由于在碱性溶液存在下BSA-AuNCs对DA的氧化和荧光猝灭,这种DA的生物传感易于实现视觉检测。比色和荧光系统对DA均表现出优于干扰物质的优异选择性。此外,我们证明了本方法在人工脑脊液(ACSF)和血清样本分析中的应用,表明该系统在诊断方面具有前景。

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