Ratiometric electrochemical proximity assay for sensitive one-step protein detection

This work proposes the concept of ratiometric electrochemical proximity assay (REPA), which can be used for one-step, highly sensitive and selective detection of protein. The assay strategy was achieved on a sensing interface that was formed by hybridization of methylene blue (MB)-labeled antibody-D...

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Autores principales: Ren, Kewei, Wu, Jie, Yan, Feng, Ju, Huangxian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950580/
https://www.ncbi.nlm.nih.gov/pubmed/24618513
http://dx.doi.org/10.1038/srep04360
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author Ren, Kewei
Wu, Jie
Yan, Feng
Ju, Huangxian
author_facet Ren, Kewei
Wu, Jie
Yan, Feng
Ju, Huangxian
author_sort Ren, Kewei
collection PubMed
description This work proposes the concept of ratiometric electrochemical proximity assay (REPA), which can be used for one-step, highly sensitive and selective detection of protein. The assay strategy was achieved on a sensing interface that was formed by hybridization of methylene blue (MB)-labeled antibody-DNA probe (MB-DNA1-Ab1) with ferrocene (Fc)-labeled DNA capture probe (Fc-P) modified gold electrode. On the interface the target protein could trigger the formation of immunocomplex between MB-DNA1-Ab1 and detection antibody-DNA probe (Ab2-DNA2) and subsequently the proximity hybridization of DNA1-DNA2, which led to the departure of MB-DNA1-Ab1 from the interface. The remained Fc-P could form a hairpin structure to take Fc group to electrode surface. Therefore, the recognition of target protein to Ab1 and Ab2 resulted in both the “signal-off” of MB and the “signal-on” of Fc for dual-signal electrochemical ratiometric readout. The proposed REPA could be carried out in one-step with 40-min duration and showed a wide detection range from 0.05 to 100 ng/mL with pg/mL limit of detection, displaying great potential for convenient point-of-care testing and commercial application.
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spelling pubmed-39505802014-03-19 Ratiometric electrochemical proximity assay for sensitive one-step protein detection Ren, Kewei Wu, Jie Yan, Feng Ju, Huangxian Sci Rep Article This work proposes the concept of ratiometric electrochemical proximity assay (REPA), which can be used for one-step, highly sensitive and selective detection of protein. The assay strategy was achieved on a sensing interface that was formed by hybridization of methylene blue (MB)-labeled antibody-DNA probe (MB-DNA1-Ab1) with ferrocene (Fc)-labeled DNA capture probe (Fc-P) modified gold electrode. On the interface the target protein could trigger the formation of immunocomplex between MB-DNA1-Ab1 and detection antibody-DNA probe (Ab2-DNA2) and subsequently the proximity hybridization of DNA1-DNA2, which led to the departure of MB-DNA1-Ab1 from the interface. The remained Fc-P could form a hairpin structure to take Fc group to electrode surface. Therefore, the recognition of target protein to Ab1 and Ab2 resulted in both the “signal-off” of MB and the “signal-on” of Fc for dual-signal electrochemical ratiometric readout. The proposed REPA could be carried out in one-step with 40-min duration and showed a wide detection range from 0.05 to 100 ng/mL with pg/mL limit of detection, displaying great potential for convenient point-of-care testing and commercial application. Nature Publishing Group 2014-03-12 /pmc/articles/PMC3950580/ /pubmed/24618513 http://dx.doi.org/10.1038/srep04360 Text en Copyright © 2014, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareALike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
spellingShingle Article
Ren, Kewei
Wu, Jie
Yan, Feng
Ju, Huangxian
Ratiometric electrochemical proximity assay for sensitive one-step protein detection
title Ratiometric electrochemical proximity assay for sensitive one-step protein detection
title_full Ratiometric electrochemical proximity assay for sensitive one-step protein detection
title_fullStr Ratiometric electrochemical proximity assay for sensitive one-step protein detection
title_full_unstemmed Ratiometric electrochemical proximity assay for sensitive one-step protein detection
title_short Ratiometric electrochemical proximity assay for sensitive one-step protein detection
title_sort ratiometric electrochemical proximity assay for sensitive one-step protein detection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950580/
https://www.ncbi.nlm.nih.gov/pubmed/24618513
http://dx.doi.org/10.1038/srep04360
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