A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic Rapamycin
An antibiotic rapamycin is one of the most commonly used immunosuppressive drugs, and also implicated for its anti-cancer activity. Hence, the determination of its blood level after organ transplantation or tumor treatment is of great concern in medicine. Although there are several rapamycin detecti...
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| Format: | Article |
| Language: | English |
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MDPI AG
2015-03-01
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| Series: | Biosensors |
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| Online Access: | http://www.mdpi.com/2079-6374/5/2/131 |
| _version_ | 1811238617466535936 |
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| author | Hee-Jin Jeong Shuya Itayama Hiroshi Ueda |
| author_facet | Hee-Jin Jeong Shuya Itayama Hiroshi Ueda |
| author_sort | Hee-Jin Jeong |
| collection | DOAJ |
| description | An antibiotic rapamycin is one of the most commonly used immunosuppressive drugs, and also implicated for its anti-cancer activity. Hence, the determination of its blood level after organ transplantation or tumor treatment is of great concern in medicine. Although there are several rapamycin detection methods, many of them have limited sensitivity, and/or need complicated procedures and long assay time. As a novel fluorescent biosensor for rapamycin, here we propose “Q’-body”, which works on the fluorescence quench-release principle inspired by the antibody-based quenchbody (Q-body) technology. We constructed rapamycin Q’-bodies by linking the two interacting domains FKBP12 and FRB, whose association is triggered by rapamycin. The fusion proteins were each incorporated position-specifically with one of fluorescence dyes ATTO520, tetramethylrhodamine, or ATTO590 using a cell-free translation system. As a result, rapid rapamycin dose-dependent fluorescence increase derived of Q’-bodies was observed, especially for those with ATTO520 with a lowest detection limit of 0.65 nM, which indicates its utility as a novel fluorescent biosensor for rapamycin. |
| first_indexed | 2024-04-12T12:44:38Z |
| format | Article |
| id | doaj.art-1ae3fabeb3af44348e14602e117744ae |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-04-12T12:44:38Z |
| publishDate | 2015-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-1ae3fabeb3af44348e14602e117744ae2022-12-22T03:32:38ZengMDPI AGBiosensors2079-63742015-03-015213114010.3390/bios5020131bios5020131A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic RapamycinHee-Jin Jeong0Shuya Itayama1Hiroshi Ueda2Chemical Resources Laboratory, Tokyo Institute of Technology, 4259-R1-18 Nagatsuta-cho, Midori-ku, Yokohama, 226-8503, JapanDepartment of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, JapanChemical Resources Laboratory, Tokyo Institute of Technology, 4259-R1-18 Nagatsuta-cho, Midori-ku, Yokohama, 226-8503, JapanAn antibiotic rapamycin is one of the most commonly used immunosuppressive drugs, and also implicated for its anti-cancer activity. Hence, the determination of its blood level after organ transplantation or tumor treatment is of great concern in medicine. Although there are several rapamycin detection methods, many of them have limited sensitivity, and/or need complicated procedures and long assay time. As a novel fluorescent biosensor for rapamycin, here we propose “Q’-body”, which works on the fluorescence quench-release principle inspired by the antibody-based quenchbody (Q-body) technology. We constructed rapamycin Q’-bodies by linking the two interacting domains FKBP12 and FRB, whose association is triggered by rapamycin. The fusion proteins were each incorporated position-specifically with one of fluorescence dyes ATTO520, tetramethylrhodamine, or ATTO590 using a cell-free translation system. As a result, rapid rapamycin dose-dependent fluorescence increase derived of Q’-bodies was observed, especially for those with ATTO520 with a lowest detection limit of 0.65 nM, which indicates its utility as a novel fluorescent biosensor for rapamycin.http://www.mdpi.com/2079-6374/5/2/131fluorescent biosensorfluorescence quenchingrapamycinantibioticsphotoinduced electron transfer |
| spellingShingle | Hee-Jin Jeong Shuya Itayama Hiroshi Ueda A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic Rapamycin Biosensors fluorescent biosensor fluorescence quenching rapamycin antibiotics photoinduced electron transfer |
| title | A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic Rapamycin |
| title_full | A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic Rapamycin |
| title_fullStr | A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic Rapamycin |
| title_full_unstemmed | A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic Rapamycin |
| title_short | A Signal-On Fluorosensor Based on Quench-Release Principle for Sensitive Detection of Antibiotic Rapamycin |
| title_sort | signal on fluorosensor based on quench release principle for sensitive detection of antibiotic rapamycin |
| topic | fluorescent biosensor fluorescence quenching rapamycin antibiotics photoinduced electron transfer |
| url | http://www.mdpi.com/2079-6374/5/2/131 |
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