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Label-Free and Substrate-Free Potentiometric Aptasensing Using Polycation-Sensitive Membrane Electrodes
Ding, Jiawang1,2; Chen, Yan1; Wang, Xuewei1; Qin, Wei1
2012-02-21
发表期刊ANALYTICAL CHEMISTRY
ISSN0003-2700
卷号84期号:4页码:2055-2061
产权排序[Ding, Jiawang; Chen, Yan; Wang, Xuewei; Qin, Wei] Chinese Acad Sci, Shandong Prov Key Lab Coastal Zone Environm Proc, Key Lab Coastal Zone Environm Proc, Yantai Inst Coastal Zone Res YIC,YICCAS, Yantai 264003, Shandong, Peoples R China; [Ding, Jiawang] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
摘要A potentiometric label-free and substrate-free (LFSF) aptasensing strategy which eliminates the labeling, separation, and immobilization steps is described in this paper. An aptamer binds specifically to a target molecule via reaction incubation, which could induce a change in the aptamer conformation from a random coil-like configuration to a rigid folded structure. Such a target binding-induced aptamer conformational change effectively prevents the aptamer from electrostatically interacting with the protamine binding domain. This could either shift the response curve for the potentiometric titration of the aptamer with protamine as monitored by a conventional polycation-sensitive membrane electrode or change the current-dependent potential detected by a protamine-conditioned polycation-sensitive electrode with the pulsed current-driven ion fluxes of protamine across the polymeric membrane. Using adenosine triphosphate (ATP) as a model analyte, the proposed concept offers potentiometric detection of ATP down to the submicromolar concentration range and has been applied to the determination of ATP in HeLa cells. In contrast to the current LFSF aptasensors based on optical detection, the proposed strategy allows the LFSF biosensing of aptamer/target binding events in a homogeneous solution via electrochemical transduction. It is anticipated that the proposed strategy will lay a foundation for development of potentiometric sensors for LFSF aptasensing of a variety of analytes where target binding-induced conformational changes such as the formation of folded structures and the opening of DNA hairpin loops are involved.; A potentiometric label-free and substrate-free (LFSF) aptasensing strategy which eliminates the labeling, separation, and immobilization steps is described in this paper. An aptamer binds specifically to a target molecule via reaction incubation, which could induce a change in the aptamer conformation from a random coil-like configuration to a rigid folded structure. Such a target binding-induced aptamer conformational change effectively prevents the aptamer from electrostatically interacting with the protamine binding domain. This could either shift the response curve for the potentiometric titration of the aptamer with protamine as monitored by a conventional polycation-sensitive membrane electrode or change the current-dependent potential detected by a protamine-conditioned polycation-sensitive electrode with the pulsed current-driven ion fluxes of protamine across the polymeric membrane. Using adenosine triphosphate (ATP) as a model analyte, the proposed concept offers potentiometric detection of ATP down to the submicromolar concentration range and has been applied to the determination of ATP in HeLa cells. In contrast to the current LFSF aptasensors based on optical detection, the proposed strategy allows the LFSF biosensing of aptamer/target binding events in a homogeneous solution via electrochemical transduction. It is anticipated that the proposed strategy will lay a foundation for development of potentiometric sensors for LFSF aptasensing of a variety of analytes where target binding-induced conformational changes such as the formation of folded structures and the opening of DNA hairpin loops are involved.
通讯作者Qin, W (reprint author), Chinese Acad Sci, Shandong Prov Key Lab Coastal Zone Environm Proc, Key Lab Coastal Zone Environm Proc, Yantai Inst Coastal Zone Res YIC,YICCAS, Yantai 264003, Shandong, Peoples R China. wqin@yic.ac.cn
关键词Aptamer-based Biosensors Polyion Sensors Electrochemical Sensor Molecular Recognition Reversible Detection Optical-detection Living Cells Dna Protamine Atp
作者部门环境化学实验室 
学科领域Chemistry
项目资助者National Natural Science Foundation of China[41176081]; Outstanding Youth Natural Science Foundation of Shandong Province[JQ200814]; Taishan Scholar Program of Shandong Province[TS20081159]
URL查看原文
收录类别SCI
关键词[WOS]APTAMER-BASED BIOSENSORS ; POLYION SENSORS ; ELECTROCHEMICAL SENSOR ; MOLECULAR RECOGNITION ; REVERSIBLE DETECTION ; OPTICAL-DETECTION ; LIVING CELLS ; DNA ; PROTAMINE ; ATP
文章类型Article
语种英语
WOS研究方向Chemistry
WOS记录号WOS:000300470800038
引用统计
文献类型期刊论文
条目标识符http://ir.yic.ac.cn/handle/133337/5510
专题环境化学实验室
作者单位1.Chinese Acad Sci, Shandong Prov Key Lab Coastal Zone Environm Proc, Key Lab Coastal Zone Environm Proc, Yantai Inst Coastal Zone Res YIC,YICCAS, Yantai 264003, Shandong, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
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GB/T 7714
Ding, Jiawang,Chen, Yan,Wang, Xuewei,et al. Label-Free and Substrate-Free Potentiometric Aptasensing Using Polycation-Sensitive Membrane Electrodes[J]. ANALYTICAL CHEMISTRY,2012,84(4):2055-2061.
APA Ding, Jiawang,Chen, Yan,Wang, Xuewei,&Qin, Wei.(2012).Label-Free and Substrate-Free Potentiometric Aptasensing Using Polycation-Sensitive Membrane Electrodes.ANALYTICAL CHEMISTRY,84(4),2055-2061.
MLA Ding, Jiawang,et al."Label-Free and Substrate-Free Potentiometric Aptasensing Using Polycation-Sensitive Membrane Electrodes".ANALYTICAL CHEMISTRY 84.4(2012):2055-2061.
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