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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| Format: | Article |
| Language: | English |
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MDPI AG
2021-10-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/11/11/422 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-10T05:39:53Z |
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| id | doaj.art-037010bdf73d47488a9cda6105cead6a |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-10T05:39:53Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-037010bdf73d47488a9cda6105cead6a2023-11-22T22:35:55ZengMDPI AGBiosensors2079-63742021-10-01111142210.3390/bios11110422DNAzyme-Amplified Label-Free Biosensor for the Simple and Sensitive Detection of PyrophosphataseCheng-Yu Lee0Chi-Hsiang Liao1Nei-Mei Fang2You-Zung Hsieh3Department of Applied Chemistry, National Yang Ming Chiao Tung University, Hsinchu 300, TaiwanDepartment of Applied Chemistry, National Yang Ming Chiao Tung University, Hsinchu 300, TaiwanDepartment of Applied Chemistry, National Yang Ming Chiao Tung University, Hsinchu 300, TaiwanDepartment of Applied Chemistry, National Yang Ming Chiao Tung University, Hsinchu 300, TaiwanThe 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.https://www.mdpi.com/2079-6374/11/11/422biosensorPPaseDNAzymeATMNDAP site |
| spellingShingle | 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 Biosensors biosensor PPase DNAzyme ATMND AP site |
| title | 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_short | 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 |
| topic | biosensor PPase DNAzyme ATMND AP site |
| url | https://www.mdpi.com/2079-6374/11/11/422 |
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