A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs.
Biosensors have emerged as a valuable tool with high specificity and sensitivity for fast and reliable detection of hazardous substances in drinking water. Numerous substances have been addressed using synthetic biology approaches. However, many proposed biosensors are based on living, genetically m...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2019-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0210940 |
| _version_ | 1819018824471543808 |
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| author | Alexander Gräwe Anna Dreyer Tobias Vornholt Ursela Barteczko Luzia Buchholz Gila Drews Uyen Linh Ho Marta Eva Jackowski Melissa Kracht Janina Lüders Tore Bleckwehl Lukas Rositzka Matthias Ruwe Manuel Wittchen Petra Lutter Kristian Müller Jörn Kalinowski |
| author_facet | Alexander Gräwe Anna Dreyer Tobias Vornholt Ursela Barteczko Luzia Buchholz Gila Drews Uyen Linh Ho Marta Eva Jackowski Melissa Kracht Janina Lüders Tore Bleckwehl Lukas Rositzka Matthias Ruwe Manuel Wittchen Petra Lutter Kristian Müller Jörn Kalinowski |
| author_sort | Alexander Gräwe |
| collection | DOAJ |
| description | Biosensors have emerged as a valuable tool with high specificity and sensitivity for fast and reliable detection of hazardous substances in drinking water. Numerous substances have been addressed using synthetic biology approaches. However, many proposed biosensors are based on living, genetically modified organisms and are therefore limited in shelf life, usability and biosafety. We addressed these issues by the construction of an extensible, cell-free biosensor. Storage is possible through freeze drying on paper. Following the addition of an aqueous sample, a highly efficient cell-free protein synthesis (CFPS) reaction is initiated. Specific allosteric transcription factors modulate the expression of 'superfolder' green fluorescent protein (sfGFP) depending on the presence of the substance of interest. The resulting fluorescence intensities are analyzed with a conventional smartphone accompanied by simple and cheap light filters. An ordinary differential equitation (ODE) model of the biosensors was developed, which enabled prediction and optimization of performance. With an optimized cell-free biosensor based on the Shigella flexneri MerR transcriptional activator, detection of 6 μg/L Hg(II) ions in water was achieved. Furthermore, a completely new biosensor for the detection of gamma-hydroxybutyrate (GHB), a substance used as date-rape drug, was established by employing the naturally occurring transcriptional repressor BlcR from Agrobacterium tumefaciens. |
| first_indexed | 2024-12-21T03:25:33Z |
| format | Article |
| id | doaj.art-ded62c19d1814ee4853fd59f84eba526 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T03:25:33Z |
| publishDate | 2019-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ded62c19d1814ee4853fd59f84eba5262022-12-21T19:17:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-01143e021094010.1371/journal.pone.0210940A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs.Alexander GräweAnna DreyerTobias VornholtUrsela BarteczkoLuzia BuchholzGila DrewsUyen Linh HoMarta Eva JackowskiMelissa KrachtJanina LüdersTore BleckwehlLukas RositzkaMatthias RuweManuel WittchenPetra LutterKristian MüllerJörn KalinowskiBiosensors have emerged as a valuable tool with high specificity and sensitivity for fast and reliable detection of hazardous substances in drinking water. Numerous substances have been addressed using synthetic biology approaches. However, many proposed biosensors are based on living, genetically modified organisms and are therefore limited in shelf life, usability and biosafety. We addressed these issues by the construction of an extensible, cell-free biosensor. Storage is possible through freeze drying on paper. Following the addition of an aqueous sample, a highly efficient cell-free protein synthesis (CFPS) reaction is initiated. Specific allosteric transcription factors modulate the expression of 'superfolder' green fluorescent protein (sfGFP) depending on the presence of the substance of interest. The resulting fluorescence intensities are analyzed with a conventional smartphone accompanied by simple and cheap light filters. An ordinary differential equitation (ODE) model of the biosensors was developed, which enabled prediction and optimization of performance. With an optimized cell-free biosensor based on the Shigella flexneri MerR transcriptional activator, detection of 6 μg/L Hg(II) ions in water was achieved. Furthermore, a completely new biosensor for the detection of gamma-hydroxybutyrate (GHB), a substance used as date-rape drug, was established by employing the naturally occurring transcriptional repressor BlcR from Agrobacterium tumefaciens.https://doi.org/10.1371/journal.pone.0210940 |
| spellingShingle | Alexander Gräwe Anna Dreyer Tobias Vornholt Ursela Barteczko Luzia Buchholz Gila Drews Uyen Linh Ho Marta Eva Jackowski Melissa Kracht Janina Lüders Tore Bleckwehl Lukas Rositzka Matthias Ruwe Manuel Wittchen Petra Lutter Kristian Müller Jörn Kalinowski A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs. PLoS ONE |
| title | A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs. |
| title_full | A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs. |
| title_fullStr | A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs. |
| title_full_unstemmed | A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs. |
| title_short | A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs. |
| title_sort | paper based cell free biosensor system for the detection of heavy metals and date rape drugs |
| url | https://doi.org/10.1371/journal.pone.0210940 |
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