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Protein sensors combining both on-and-off model for antibody homogeneous assay.
Li, Jie; Wang, Jin-Lan; Zhang, Wen-Lu; Tu, Zeng; Cai, Xue-Fei; Wang, Yu-Wei; Gan, Chun-Yang; Deng, Hai-Jun; Cui, Jing; Shu, Zhao-Che; Long, Quan-Xin; Chen, Juan; Tang, Ni; Hu, Xue; Huang, Ai-Long; Hu, Jie-Li.
  • Li J; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Wang JL; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Zhang WL; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Tu Z; Department of Immunology and Microbiology, Chongqing Medical University, Chongqing, China.
  • Cai XF; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Wang YW; Department of Laboratory Medicine, Chongqing Hospital of Traditional Chinese Medicine, China; Laboratory for Diagnosis and Treatment of Infectious Diseases Integrated Traditional Chinese and Western Medicine, Chongqing Hospital of Traditional Chinese Medicine, China.
  • Gan CY; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Deng HJ; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Cui J; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Shu ZC; The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
  • Long QX; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Chen J; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Tang N; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Hu X; The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China. Electronic address: huxue@hospital.cqmu.edu.cn.
  • Huang AL; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China. Electronic address: ahuang@cqmu.edu.cn.
  • Hu JL; Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China. Electronic address: 102564@cqmu.edu.cn.
Biosens Bioelectron ; 209: 114226, 2022 Aug 01.
Article in English | MEDLINE | ID: covidwho-1767929
ABSTRACT
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Biosens Bioelectron Journal subject: Biotechnology Year: 2022 Document Type: Article Affiliation country: J.bios.2022.114226

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Biosens Bioelectron Journal subject: Biotechnology Year: 2022 Document Type: Article Affiliation country: J.bios.2022.114226