ABSTRACT
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is currently spreading and mutating with increasing speed worldwide. Therefore, there is an urgent need for a simple, sensitive, and high-throughput (HTP) assay to quantify virus-host interaction in order to quickly evaluate infectious ability of mutant virus and develop or validate virus-inhibiting drugs. Here we have developed an ultrasensitive bioluminescent biosensor to evaluate virus-cell interaction by quantifying the interaction between SARS-CoV-2 receptor binding domain (RBD) and its cellular receptor angiotensin-converting enzyme 2 (ACE2) both in living cells and in vitro. We have successfully used this novel biosensor to analyze SARS-CoV-2 RBD mutants, and evaluated candidate small molecules (SMs), antibodies, and peptides that may block RBD:ACE2 interaction. This simple, rapid and HTP biosensor tool will significantly expedite detection of viral mutants and anti-COVID-19 drug discovery processes.
Competing Interest Statement
The authors have declared no competing interest.
ABBREVIATIONS
- aa
- amino acid
- ACE2
- angiotensin-converting enzyme 2
- DMEM
- Dulbecco’s modified Eagle’s medium
- HTP
- high throughput
- Lg/LgBiT
- large BiT
- RBD
- receptor binding domain
- PAGE
- polyacrylamide gel electrophoresis
- S
- spike
- SARS
- severe acute respiratory syndrome coronavirus-2
- SM
- small molecule
- Sm/SmBiT
- small BiT
- SPR
- surface plasmon resonance
- SLCA
- split luciferase complementary assay
- WT
- wild-type