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Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor
The SELEX (Systematic Evolution of Ligands by EXponential enrichment) method has been used successfully since 1990, but work is still required to obtain highly specific aptamers. Here, we present a novel approach called ‘Competitive non-SELEX’ (and termed as ‘SELCOS’ (Systematic Evolution of Ligands...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6491428/ https://www.ncbi.nlm.nih.gov/pubmed/31040350 http://dx.doi.org/10.1038/s41598-019-43187-6 |
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author | Kushwaha, Ankita Takamura, Yuzuru Nishigaki, Koichi Biyani, Manish |
author_facet | Kushwaha, Ankita Takamura, Yuzuru Nishigaki, Koichi Biyani, Manish |
author_sort | Kushwaha, Ankita |
collection | PubMed |
description | The SELEX (Systematic Evolution of Ligands by EXponential enrichment) method has been used successfully since 1990, but work is still required to obtain highly specific aptamers. Here, we present a novel approach called ‘Competitive non-SELEX’ (and termed as ‘SELCOS’ (Systematic Evolution of Ligands by COmpetitive Selection)) for readily obtaining aptamers that can discriminate between highly similar targets. This approach is based on the theoretical background presented here, in which under the co-presence of two similar targets, a specific binding type can be enriched more than a nonspecifically binding one during repetitive steps of partitioning with no PCR amplification between them. This principle was experimentally confirmed by the selection experiment for influenza virus subtype-specific DNA aptamers. Namely, the selection products (pools of DNA aptamers) obtained by SELCOS were subjected to a DEPSOR-mode electrochemical sensor, enabling the method to select subtype-specific aptamer pools. From the clonal analysis of these pools, only a few rounds of in vitro selection were sufficient to achieve the surprisingly rapid enrichment of a small number of aptamers with high selectivity, which could be attributed to the SELCOS principle and the given selection pressure program. The subtype-specific aptamers obtained in this manner had a high affinity (e.g., K(D) = 82 pM for H1N1; 88 pM for H3N2) and negligible cross-reactivity. By making the H1N1-specific DNA aptamer a sensor unit of the DEPSOR electrochemical detector, an influenza virus subtype-specific and portable detector was readily constructed, indicating how close it is to the field application goal. |
format | Online Article Text |
id | pubmed-6491428 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-64914282019-05-16 Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor Kushwaha, Ankita Takamura, Yuzuru Nishigaki, Koichi Biyani, Manish Sci Rep Article The SELEX (Systematic Evolution of Ligands by EXponential enrichment) method has been used successfully since 1990, but work is still required to obtain highly specific aptamers. Here, we present a novel approach called ‘Competitive non-SELEX’ (and termed as ‘SELCOS’ (Systematic Evolution of Ligands by COmpetitive Selection)) for readily obtaining aptamers that can discriminate between highly similar targets. This approach is based on the theoretical background presented here, in which under the co-presence of two similar targets, a specific binding type can be enriched more than a nonspecifically binding one during repetitive steps of partitioning with no PCR amplification between them. This principle was experimentally confirmed by the selection experiment for influenza virus subtype-specific DNA aptamers. Namely, the selection products (pools of DNA aptamers) obtained by SELCOS were subjected to a DEPSOR-mode electrochemical sensor, enabling the method to select subtype-specific aptamer pools. From the clonal analysis of these pools, only a few rounds of in vitro selection were sufficient to achieve the surprisingly rapid enrichment of a small number of aptamers with high selectivity, which could be attributed to the SELCOS principle and the given selection pressure program. The subtype-specific aptamers obtained in this manner had a high affinity (e.g., K(D) = 82 pM for H1N1; 88 pM for H3N2) and negligible cross-reactivity. By making the H1N1-specific DNA aptamer a sensor unit of the DEPSOR electrochemical detector, an influenza virus subtype-specific and portable detector was readily constructed, indicating how close it is to the field application goal. Nature Publishing Group UK 2019-04-30 /pmc/articles/PMC6491428/ /pubmed/31040350 http://dx.doi.org/10.1038/s41598-019-43187-6 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Kushwaha, Ankita Takamura, Yuzuru Nishigaki, Koichi Biyani, Manish Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor |
title | Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor |
title_full | Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor |
title_fullStr | Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor |
title_full_unstemmed | Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor |
title_short | Competitive non-SELEX for the selective and rapid enrichment of DNA aptamers and its use in electrochemical aptasensor |
title_sort | competitive non-selex for the selective and rapid enrichment of dna aptamers and its use in electrochemical aptasensor |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6491428/ https://www.ncbi.nlm.nih.gov/pubmed/31040350 http://dx.doi.org/10.1038/s41598-019-43187-6 |
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