Selection, Characterization, and Application of ssDNA Aptamer against Furaneol
Furaneol is an aroma compound which occurs naturally in foods and is used as an artificial flavor. Detection of furaneol is required in food science and food processing industry. Capture- Systematic Evolution of Ligands by EXponential enrichment (SELEX) protocol was applied for the isolation of an a...
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| Format: | Article |
| Language: | English |
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MDPI AG
2018-11-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/23/12/3159 |
| _version_ | 1818991399727529984 |
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| author | Natalia Komarova Mariia Andrianova Sergey Glukhov Alexander Kuznetsov |
| author_facet | Natalia Komarova Mariia Andrianova Sergey Glukhov Alexander Kuznetsov |
| author_sort | Natalia Komarova |
| collection | DOAJ |
| description | Furaneol is an aroma compound which occurs naturally in foods and is used as an artificial flavor. Detection of furaneol is required in food science and food processing industry. Capture- Systematic Evolution of Ligands by EXponential enrichment (SELEX) protocol was applied for the isolation of an aptamer binding to furaneol, a small volatile organic substance contributing to the flavor of various products. Thirteen cycles of selection were performed. The resulting DNA pool was cloned, using blunt-end cloning, and ninety-six plasmids were sequenced and analyzed. Eight oligonucleotides were selected as aptamer candidates and screened for the ability to bind to furaneol, using three different methods—magnetic-beads associated elution assay, SYBR Green I assay, and exonuclease protection assay. One of the candidates was further characterized as an aptamer. The apparent equilibrium constant was determined to be (1.1 ± 0.4) µM, by the fluorescent method. The reported aptamer was applied for development of the ion-sensitive field-effect transistor (ISFET)-based biosensor, for the analysis of furaneol, in the concentration range of 0.1⁻10 µM. |
| first_indexed | 2024-12-20T20:09:39Z |
| format | Article |
| id | doaj.art-1e4853d391774bad8e9cb7814ebe03df |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-20T20:09:39Z |
| publishDate | 2018-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-1e4853d391774bad8e9cb7814ebe03df2022-12-21T19:27:50ZengMDPI AGMolecules1420-30492018-11-012312315910.3390/molecules23123159molecules23123159Selection, Characterization, and Application of ssDNA Aptamer against FuraneolNatalia Komarova0Mariia Andrianova1Sergey Glukhov2Alexander Kuznetsov3Scientific-Manufacturing Complex Technological Centre, 1–7 Shokin Square, Zelenograd, 124498 Moscow, RussiaScientific-Manufacturing Complex Technological Centre, 1–7 Shokin Square, Zelenograd, 124498 Moscow, RussiaScientific-Manufacturing Complex Technological Centre, 1–7 Shokin Square, Zelenograd, 124498 Moscow, RussiaScientific-Manufacturing Complex Technological Centre, 1–7 Shokin Square, Zelenograd, 124498 Moscow, RussiaFuraneol is an aroma compound which occurs naturally in foods and is used as an artificial flavor. Detection of furaneol is required in food science and food processing industry. Capture- Systematic Evolution of Ligands by EXponential enrichment (SELEX) protocol was applied for the isolation of an aptamer binding to furaneol, a small volatile organic substance contributing to the flavor of various products. Thirteen cycles of selection were performed. The resulting DNA pool was cloned, using blunt-end cloning, and ninety-six plasmids were sequenced and analyzed. Eight oligonucleotides were selected as aptamer candidates and screened for the ability to bind to furaneol, using three different methods—magnetic-beads associated elution assay, SYBR Green I assay, and exonuclease protection assay. One of the candidates was further characterized as an aptamer. The apparent equilibrium constant was determined to be (1.1 ± 0.4) µM, by the fluorescent method. The reported aptamer was applied for development of the ion-sensitive field-effect transistor (ISFET)-based biosensor, for the analysis of furaneol, in the concentration range of 0.1⁻10 µM.https://www.mdpi.com/1420-3049/23/12/3159aptamerfuraneolCapture-SELEXISFETaptasensor |
| spellingShingle | Natalia Komarova Mariia Andrianova Sergey Glukhov Alexander Kuznetsov Selection, Characterization, and Application of ssDNA Aptamer against Furaneol Molecules aptamer furaneol Capture-SELEX ISFET aptasensor |
| title | Selection, Characterization, and Application of ssDNA Aptamer against Furaneol |
| title_full | Selection, Characterization, and Application of ssDNA Aptamer against Furaneol |
| title_fullStr | Selection, Characterization, and Application of ssDNA Aptamer against Furaneol |
| title_full_unstemmed | Selection, Characterization, and Application of ssDNA Aptamer against Furaneol |
| title_short | Selection, Characterization, and Application of ssDNA Aptamer against Furaneol |
| title_sort | selection characterization and application of ssdna aptamer against furaneol |
| topic | aptamer furaneol Capture-SELEX ISFET aptasensor |
| url | https://www.mdpi.com/1420-3049/23/12/3159 |
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