| Autor: |
Jie Li, Jin-Lan Wang, Wen-Lu Zhang, Zeng Tu, Xue-Fei Cai, Yu-Wei Wang, Chun-Yang Gan, Hai-Jun Deng, Jing Cui, Zhao-Che Shu, Quan-Xin Long, Juan Chen, Ni Tang, Xue Hu, Ai-Long Huang, Jie-Li Hu |
| Rok vydání: |
2021 |
| Předmět: |
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| Zdroj: |
Biosensorsbioelectronics. 209 |
| ISSN: |
1873-4235 |
| Popis: |
Protein sensors based on allosteric enzymes responding to target binding with rapid changes in enzymatic activity are potential tools for homogeneous assays. However, a high signal-to-noise ratio (S/N) is difficult to achieve in their construction. A high S/N is critical to discriminate signals from the background, a phenomenon that might largely vary among serum samples from different individuals. Herein, based on the modularized luciferase NanoLuc, we designed a novel biosensor called NanoSwitch. This sensor allows direct detection of antibodies in 1 μl serum in 45 min without washing steps. In the detection of Flag and HA antibodies, NanoSwitches respond to antibodies with S/N ratios of 33-fold and 42-fold, respectively. Further, we constructed a NanoSwitch for detecting SARS-CoV-2-specific antibodies, which showed over 200-fold S/N in serum samples. High S/N was achieved by a new working model, combining the turn-off of the sensor with human serum albumin and turn-on with a specific antibody. Also, we constructed NanoSwitches for detecting antibodies against the core protein of hepatitis C virus (HCV) and gp41 of the human immunodeficiency virus (HIV). Interestingly, these sensors demonstrated a high S/N and good performance in the assays of clinical samples; this was partly attributed to the combination of off-and-on models. In summary, we provide a novel type of protein sensor and a working model that potentially guides new sensor design with better performance. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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