Bioluminescent Protein–Inhibitor Pair in the Design of a Molecular Aptamer Beacon Biosensing System
| العنوان: | Bioluminescent Protein–Inhibitor Pair in the Design of a Molecular Aptamer Beacon Biosensing System |
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| المؤلفون: | Angel E. Kaifer, Angeliki Moutsiopoulou, David Broyles, Emre Dikici, Hamdi Joda, Sylvia Daunert, Avinash Kaur, Sapna K. Deo |
| المصدر: | Anal Chem |
| بيانات النشر: | American Chemical Society (ACS), 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Models, Molecular, Aptamer, Biosensing Techniques, Signal-To-Noise Ratio, 010402 general chemistry, 01 natural sciences, Article, Analytical Chemistry, Copepoda, chemistry.chemical_compound, Gaussia, Molecular beacon, Coelenterazine, Animals, Bioluminescence, Luciferase, Enzyme Inhibitors, Luciferases, Quenching (fluorescence), Molecular Structure, biology, 010401 analytical chemistry, Imidazoles, Aptamers, Nucleotide, biology.organism_classification, 0104 chemical sciences, Luminescent Proteins, chemistry, Pyrazines, Luminescent Measurements, Biophysics, Biosensor |
| الوصف: | Although bioluminescent molecular beacons designed around resonance quenchers have shown higher signal-to-noise ratios and increased sensitivity compared with fluorescent beacon systems, bioluminescence quenching is still comparatively inefficient. A more elegant solution to inefficient quenching can be realized by designing a competitive inhibitor that is structurally very similar to the native substrate, resulting in essentially complete substrate exclusion. In this work, we designed a conjugated anti-interferon-γ (IFN-γ) molecular aptamer beacon (MAB) attached to a bioluminescent protein, Gaussia luciferase (GLuc), and an inhibitor molecule with a similar structure to the native substrate coelenterazine. To prove that a MAB can be more sensitive and have a better signal-to-noise ratio, a bioluminescence-based assay was developed against IFN-γ and provided an optimized, physiologically relevant detection limit of 1.0 nM. We believe that this inhibitor approach may provide a simple alternative strategy to standard resonance quenching in the development of high-performance molecular beacon-based biosensing systems. |
| تدمد: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.0c00518 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::202cf2dc1b69362de0b7651fc7a585d9 https://doi.org/10.1021/acs.analchem.0c00518 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....202cf2dc1b69362de0b7651fc7a585d9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15206882 00032700 |
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| DOI: | 10.1021/acs.analchem.0c00518 |