A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin

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...

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Autores principales:	Xinling Zeng, Qing Zhou, Liyan Wang, Xiaoxian Zhu, Kuiyan Cui, Xinsheng Peng, Terry W. J. Steele, Huizhi Chen, Hui Xu, Yubin Zhou
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	thrombin aptamer biosensors photoisomerization fluorescence kinetic mode Technology T Electrical engineering. Electronics. Nuclear engineering TK1-9971 Engineering (General). Civil engineering (General) TA1-2040 Microscopy QH201-278.5 Descriptive and experimental mechanics QC120-168.85
Acceso en línea:	https://doaj.org/article/5496855410164ca6998117d16c1c1607
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spelling	oai:doaj.org-article:5496855410164ca6998117d16c1c16072021-11-25T18:14:46ZA Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin10.3390/ma142269271996-1944https://doaj.org/article/5496855410164ca6998117d16c1c16072021-11-01T00:00:00Zhttps://www.mdpi.com/1996-1944/14/22/6927https://doaj.org/toc/1996-1944It 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 (<i>k</i><sub>app</sub>), 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 <i>k</i><sub>app</sub> 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.Xinling ZengQing ZhouLiyan WangXiaoxian ZhuKuiyan CuiXinsheng PengTerry W. J. SteeleHuizhi ChenHui XuYubin ZhouMDPI AGarticlethrombinaptamerbiosensorsphotoisomerizationfluorescencekinetic modeTechnologyTElectrical engineering. Electronics. Nuclear engineeringTK1-9971Engineering (General). Civil engineering (General)TA1-2040MicroscopyQH201-278.5Descriptive and experimental mechanicsQC120-168.85ENMaterials, Vol 14, Iss 6927, p 6927 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	thrombin aptamer biosensors photoisomerization fluorescence kinetic mode Technology T Electrical engineering. Electronics. Nuclear engineering TK1-9971 Engineering (General). Civil engineering (General) TA1-2040 Microscopy QH201-278.5 Descriptive and experimental mechanics QC120-168.85
spellingShingle	thrombin aptamer biosensors photoisomerization fluorescence kinetic mode Technology T Electrical engineering. Electronics. Nuclear engineering TK1-9971 Engineering (General). Civil engineering (General) TA1-2040 Microscopy QH201-278.5 Descriptive and experimental mechanics QC120-168.85 Xinling Zeng Qing Zhou Liyan Wang Xiaoxian Zhu Kuiyan Cui Xinsheng Peng Terry W. J. Steele Huizhi Chen Hui Xu Yubin Zhou A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin
description	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 (<i>k</i><sub>app</sub>), 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 <i>k</i><sub>app</sub> 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.
format	article
author	Xinling Zeng Qing Zhou Liyan Wang Xiaoxian Zhu Kuiyan Cui Xinsheng Peng Terry W. J. Steele Huizhi Chen Hui Xu Yubin Zhou
author_facet	Xinling Zeng Qing Zhou Liyan Wang Xiaoxian Zhu Kuiyan Cui Xinsheng Peng Terry W. J. Steele Huizhi Chen Hui Xu Yubin Zhou
author_sort	Xinling Zeng
title	A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin
title_short	A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin
title_full	A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin
title_fullStr	A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin
title_full_unstemmed	A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin
title_sort	fluorescence kinetic-based aptasensor employing stilbene isomerization for detection of thrombin
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/5496855410164ca6998117d16c1c1607
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A Fluorescence Kinetic-Based Aptasensor Employing Stilbene Isomerization for Detection of Thrombin

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