Cargando…
A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection
The qualitative and quantitative determination of marker protein is of great significance in the life sciences and in medicine. Here, we developed an electrochemical DNA biosensor for protein detection based on DNA self-assembly and the terminal protecting effects of small-molecule-linked DNA. This...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8615823/ https://www.ncbi.nlm.nih.gov/pubmed/34821667 http://dx.doi.org/10.3390/bios11110451 |
_version_ | 1784604198781845504 |
---|---|
author | Ouyang, Ping Fang, Chenxin Han, Jialun Zhang, Jingjing Yang, Yuxing Qing, Yang Chen, Yubing Shang, Wenyan Du, Jie |
author_facet | Ouyang, Ping Fang, Chenxin Han, Jialun Zhang, Jingjing Yang, Yuxing Qing, Yang Chen, Yubing Shang, Wenyan Du, Jie |
author_sort | Ouyang, Ping |
collection | PubMed |
description | The qualitative and quantitative determination of marker protein is of great significance in the life sciences and in medicine. Here, we developed an electrochemical DNA biosensor for protein detection based on DNA self-assembly and the terminal protecting effects of small-molecule-linked DNA. This strategy is demonstrated using the small molecule biotin and its receptor protein streptavidin (SA). We immobilized DNA with a designed structure and sequence on the surface of the gold electrode, and we named it M1-Biotin DNA. M1-Biotin DNA selectively combines with SA to generate M1-Biotin-SA DNA and protects M1-Biotin DNA from digestion by EXO III; therefore, M1-Biotin DNA remains intact on the electrode surface. M1-Biotin-SA DNA was modified with methylene blue (MB); the MB reporter molecule is located near the surface of the gold electrode, which generates a substantial electrochemical signal during the detection of SA. Through this strategy, we can exploit the presence or absence of an electrochemical signal to provide qualitative target protein determination as well as the strength of the electrochemical signal to quantitatively analyze the target protein concentration. This strategy has been proven to be used for the quantitative analysis of the interaction between biotin and streptavidin (SA). Under optimal conditions, the detection limit of the proposed biosensor is as low as 18.8 pM, and the linear range is from 0.5 nM to 5 μM, showing high sensitivity. The detection ability of this DNA biosensor in complex serum samples has also been studied. At the same time, we detected the folate receptor (FR) to confirm that this strategy can be used to detect other proteins. Therefore, this electrochemical DNA biosensor provides a sensitive, low-cost, and fast target protein detection platform, which may provide a reliable and powerful tool for early disease diagnosis. |
format | Online Article Text |
id | pubmed-8615823 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-86158232021-11-26 A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection Ouyang, Ping Fang, Chenxin Han, Jialun Zhang, Jingjing Yang, Yuxing Qing, Yang Chen, Yubing Shang, Wenyan Du, Jie Biosensors (Basel) Article The qualitative and quantitative determination of marker protein is of great significance in the life sciences and in medicine. Here, we developed an electrochemical DNA biosensor for protein detection based on DNA self-assembly and the terminal protecting effects of small-molecule-linked DNA. This strategy is demonstrated using the small molecule biotin and its receptor protein streptavidin (SA). We immobilized DNA with a designed structure and sequence on the surface of the gold electrode, and we named it M1-Biotin DNA. M1-Biotin DNA selectively combines with SA to generate M1-Biotin-SA DNA and protects M1-Biotin DNA from digestion by EXO III; therefore, M1-Biotin DNA remains intact on the electrode surface. M1-Biotin-SA DNA was modified with methylene blue (MB); the MB reporter molecule is located near the surface of the gold electrode, which generates a substantial electrochemical signal during the detection of SA. Through this strategy, we can exploit the presence or absence of an electrochemical signal to provide qualitative target protein determination as well as the strength of the electrochemical signal to quantitatively analyze the target protein concentration. This strategy has been proven to be used for the quantitative analysis of the interaction between biotin and streptavidin (SA). Under optimal conditions, the detection limit of the proposed biosensor is as low as 18.8 pM, and the linear range is from 0.5 nM to 5 μM, showing high sensitivity. The detection ability of this DNA biosensor in complex serum samples has also been studied. At the same time, we detected the folate receptor (FR) to confirm that this strategy can be used to detect other proteins. Therefore, this electrochemical DNA biosensor provides a sensitive, low-cost, and fast target protein detection platform, which may provide a reliable and powerful tool for early disease diagnosis. MDPI 2021-11-13 /pmc/articles/PMC8615823/ /pubmed/34821667 http://dx.doi.org/10.3390/bios11110451 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ouyang, Ping Fang, Chenxin Han, Jialun Zhang, Jingjing Yang, Yuxing Qing, Yang Chen, Yubing Shang, Wenyan Du, Jie A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection |
title | A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection |
title_full | A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection |
title_fullStr | A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection |
title_full_unstemmed | A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection |
title_short | A DNA Electrochemical Sensor via Terminal Protection of Small-Molecule-Linked DNA for Highly Sensitive Protein Detection |
title_sort | dna electrochemical sensor via terminal protection of small-molecule-linked dna for highly sensitive protein detection |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8615823/ https://www.ncbi.nlm.nih.gov/pubmed/34821667 http://dx.doi.org/10.3390/bios11110451 |
work_keys_str_mv | AT ouyangping adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT fangchenxin adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT hanjialun adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT zhangjingjing adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT yangyuxing adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT qingyang adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT chenyubing adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT shangwenyan adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT dujie adnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT ouyangping dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT fangchenxin dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT hanjialun dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT zhangjingjing dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT yangyuxing dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT qingyang dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT chenyubing dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT shangwenyan dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection AT dujie dnaelectrochemicalsensorviaterminalprotectionofsmallmoleculelinkeddnaforhighlysensitiveproteindetection |