| Autor: |
Alexander Gräwe, Maarten Merkx |
| Přispěvatelé: |
Immunoengineering, Protein Engineering, ICMS Core |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
ACS Sensors, 7(12), 3800-3808. American Chemical Society |
| ISSN: |
2379-3694 |
| DOI: |
10.1021/acssensors.2c01726 |
| Popis: |
Bioluminescent sensor proteins have recently gained popularity in both basic research and point-of-care diagnostics. Sensor proteins based on intramolecular complementation of split NanoLuc are particularly attractive because their intrinsic modular design enables for systematic tuning of sensor properties. Here we show how the sensitivity of these sensors can be enhanced by the introduction of catalytically inactive variants of the small SmBiT subunit (DarkBiTs) as intramolecular inhibitors. Starting from previously developed bioluminescent antibody sensor proteins (LUMABS), we developed single component, biomolecular switches with a strongly reduced background signal for the detection of three clinically relevant antibodies, anti-HIV1-p17, cetuximab (CTX), and an RSV neutralizing antibody (101F). These new dark-LUMABS sensors showed 5-13-fold increases in sensitivity which translated into lower limits of detection. The use of DarkBiTs as competitive intramolecular inhibitor domains is not limited to the LUMABS sensor family and might be used to boost the performance of other bioluminescent sensor proteins based on split luciferase complementation. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
