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Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection

In this study, we fabricated a novel electrochemical biosensing platform on the basis of target-triggered proximity hybridization-mediated isothermal exponential amplification reaction (EXPAR) for ultrasensitive protein analysis. Through rational design, the aptamers for protein recognition were int...

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Autores principales: Yu, Yanyan, Su, Gaoxing, Zhu, Hongyan, Zhu, Qing, Chen, Yong, Xu, Bohui, Li, Yuqin, Zhang, Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove Medical Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5566414/
https://www.ncbi.nlm.nih.gov/pubmed/28860756
http://dx.doi.org/10.2147/IJN.S142015
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author Yu, Yanyan
Su, Gaoxing
Zhu, Hongyan
Zhu, Qing
Chen, Yong
Xu, Bohui
Li, Yuqin
Zhang, Wei
author_facet Yu, Yanyan
Su, Gaoxing
Zhu, Hongyan
Zhu, Qing
Chen, Yong
Xu, Bohui
Li, Yuqin
Zhang, Wei
author_sort Yu, Yanyan
collection PubMed
description In this study, we fabricated a novel electrochemical biosensing platform on the basis of target-triggered proximity hybridization-mediated isothermal exponential amplification reaction (EXPAR) for ultrasensitive protein analysis. Through rational design, the aptamers for protein recognition were integrated within two DNA probes. Via proximity hybridization principle, the affinity protein-binding event was converted into DNA assembly process. The recognition of protein by aptamers can trigger the strand displacement through the increase of the local concentrations of the involved probes. As a consequence, the output DNA was displaced, which can hybridize with the duplex probes immobilized on the electrode surface subsequently, leading to the initiation of the EXPAR as well as the cleavage of duplex probes. Each cleavage will release the gold nanoparticles (AuNPs) binding sequence. With the modification of G-quadruplex sequence, electrochemical signals were yielded by the AuNPs through oxidizing 3,3′,5,5′-tetramethylbenzidine in the presence of H(2)O(2). The study we proposed exhibited high sensitivity toward platelet-derived growth factor BB (PDGF-BB) with the detection limit of 52 fM. And, this method also showed great selectivity among the PDGF isoforms and performed well in spiked human serum samples.
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spelling pubmed-55664142017-08-31 Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection Yu, Yanyan Su, Gaoxing Zhu, Hongyan Zhu, Qing Chen, Yong Xu, Bohui Li, Yuqin Zhang, Wei Int J Nanomedicine Original Research In this study, we fabricated a novel electrochemical biosensing platform on the basis of target-triggered proximity hybridization-mediated isothermal exponential amplification reaction (EXPAR) for ultrasensitive protein analysis. Through rational design, the aptamers for protein recognition were integrated within two DNA probes. Via proximity hybridization principle, the affinity protein-binding event was converted into DNA assembly process. The recognition of protein by aptamers can trigger the strand displacement through the increase of the local concentrations of the involved probes. As a consequence, the output DNA was displaced, which can hybridize with the duplex probes immobilized on the electrode surface subsequently, leading to the initiation of the EXPAR as well as the cleavage of duplex probes. Each cleavage will release the gold nanoparticles (AuNPs) binding sequence. With the modification of G-quadruplex sequence, electrochemical signals were yielded by the AuNPs through oxidizing 3,3′,5,5′-tetramethylbenzidine in the presence of H(2)O(2). The study we proposed exhibited high sensitivity toward platelet-derived growth factor BB (PDGF-BB) with the detection limit of 52 fM. And, this method also showed great selectivity among the PDGF isoforms and performed well in spiked human serum samples. Dove Medical Press 2017-08-17 /pmc/articles/PMC5566414/ /pubmed/28860756 http://dx.doi.org/10.2147/IJN.S142015 Text en © 2017 Yu et al. This work is published and licensed by Dove Medical Press Limited The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.
spellingShingle Original Research
Yu, Yanyan
Su, Gaoxing
Zhu, Hongyan
Zhu, Qing
Chen, Yong
Xu, Bohui
Li, Yuqin
Zhang, Wei
Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection
title Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection
title_full Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection
title_fullStr Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection
title_full_unstemmed Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection
title_short Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection
title_sort proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5566414/
https://www.ncbi.nlm.nih.gov/pubmed/28860756
http://dx.doi.org/10.2147/IJN.S142015
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