DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase

The level of pyrophosphatase (PPase) expression has been suggested as a potential biomarker of various cancers, and its prognostic value has been evaluated in patients suffering from lung cancer, colorectal cancer, and hyperthyroidism. However, the detection of PPase usually needs specific materials...

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Autores principales:	Cheng-Yu Lee, Chi-Hsiang Liao, Nei-Mei Fang, You-Zung Hsieh
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	biosensor PPase DNAzyme ATMND AP site Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/037010bdf73d47488a9cda6105cead6a
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spelling	oai:doaj.org-article:037010bdf73d47488a9cda6105cead6a2021-11-25T16:55:00ZDNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase10.3390/bios111104222079-6374https://doaj.org/article/037010bdf73d47488a9cda6105cead6a2021-10-01T00:00:00Zhttps://www.mdpi.com/2079-6374/11/11/422https://doaj.org/toc/2079-6374The level of pyrophosphatase (PPase) expression has been suggested as a potential biomarker of various cancers, and its prognostic value has been evaluated in patients suffering from lung cancer, colorectal cancer, and hyperthyroidism. However, the detection of PPase usually needs specific materials that require complicated, time-consuming reactions with restricted linear range and sensitivity, limiting their application in early clinical diagnosis. Herein, we developed a DNAzyme-based biosensor for the detection of PPase. In the presence of PPase, pyrophosphate (PPi) and Cu<sup>2+</sup> ions released from the PPi–Cu<sup>2+</sup>–PPi complex induce the cleavage of the DNAzyme and the corresponding substrate. An apurinic/apyrimidinic (AP) site was elaborately designed within substrates that could encase the fluorophore 2-amino-5,6,7-trimethyl-1,8-naphthyridine (ATMND). The fluorescence of ATMND was initially quenched but restored when the DNAzyme/substrate complex was hydrolyzed with the release of ATMND. In this way, the PPase activity can be estimated by detecting the increased fluorescence of the released ATMND. Under optimized conditions, the activity of PPase could be analyzed at concentrations from 0.5 to 1000 mU, with the lowest detectable concentration being 0.5 mU. This work lays a foundation for developing a DNAzyme-amplified fluorescent biosensor with a high sensitivity, a wide linear range, and single-step operation for use as an easy diagnostic for PPase analysis.Cheng-Yu LeeChi-Hsiang LiaoNei-Mei FangYou-Zung HsiehMDPI AGarticlebiosensorPPaseDNAzymeATMNDAP siteBiotechnologyTP248.13-248.65ENBiosensors, Vol 11, Iss 422, p 422 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	biosensor PPase DNAzyme ATMND AP site Biotechnology TP248.13-248.65
spellingShingle	biosensor PPase DNAzyme ATMND AP site Biotechnology TP248.13-248.65 Cheng-Yu Lee Chi-Hsiang Liao Nei-Mei Fang You-Zung Hsieh DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase
description	The level of pyrophosphatase (PPase) expression has been suggested as a potential biomarker of various cancers, and its prognostic value has been evaluated in patients suffering from lung cancer, colorectal cancer, and hyperthyroidism. However, the detection of PPase usually needs specific materials that require complicated, time-consuming reactions with restricted linear range and sensitivity, limiting their application in early clinical diagnosis. Herein, we developed a DNAzyme-based biosensor for the detection of PPase. In the presence of PPase, pyrophosphate (PPi) and Cu<sup>2+</sup> ions released from the PPi–Cu<sup>2+</sup>–PPi complex induce the cleavage of the DNAzyme and the corresponding substrate. An apurinic/apyrimidinic (AP) site was elaborately designed within substrates that could encase the fluorophore 2-amino-5,6,7-trimethyl-1,8-naphthyridine (ATMND). The fluorescence of ATMND was initially quenched but restored when the DNAzyme/substrate complex was hydrolyzed with the release of ATMND. In this way, the PPase activity can be estimated by detecting the increased fluorescence of the released ATMND. Under optimized conditions, the activity of PPase could be analyzed at concentrations from 0.5 to 1000 mU, with the lowest detectable concentration being 0.5 mU. This work lays a foundation for developing a DNAzyme-amplified fluorescent biosensor with a high sensitivity, a wide linear range, and single-step operation for use as an easy diagnostic for PPase analysis.
format	article
author	Cheng-Yu Lee Chi-Hsiang Liao Nei-Mei Fang You-Zung Hsieh
author_facet	Cheng-Yu Lee Chi-Hsiang Liao Nei-Mei Fang You-Zung Hsieh
author_sort	Cheng-Yu Lee
title	DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase
title_short	DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase
title_full	DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase
title_fullStr	DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase
title_full_unstemmed	DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase
title_sort	dnazyme-amplified label-free biosensor for the simple and sensitive detection of pyrophosphatase
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/037010bdf73d47488a9cda6105cead6a
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DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of Pyrophosphatase

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