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...
Autores principales: | , , , |
---|---|
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 |
_version_ | 1782307010368241664 |
---|---|
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. |
format | Online Article Text |
id | pubmed-3950580 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT renkewei ratiometricelectrochemicalproximityassayforsensitiveonestepproteindetection AT wujie ratiometricelectrochemicalproximityassayforsensitiveonestepproteindetection AT yanfeng ratiometricelectrochemicalproximityassayforsensitiveonestepproteindetection AT juhuangxian ratiometricelectrochemicalproximityassayforsensitiveonestepproteindetection |