Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters the cells through the binding of its spike protein (S-protein) to the cell surface-expressing angiotensin-converting enzyme 2 (ACE2). Thus, inhibition of S-protein-ACE2 binding may impede SARS-CoV-2 cell entry and attenuate the progression of Coronavirus disease 2019 (COVID-19). In this study, an electrochemical impedance spectroscopy-based biosensing platform consisting of a recombinant ACE2-coated palladium nano-thin-film electrode as the core sensing element was fabricated for the screening of potential inhibitors against S-protein-ACE2 binding. The platform could detect interference of small analytes against S-protein-ACE2 binding at low analyte concentration and small volume (0.1 μg/mL and ~1 μL, estimated total analyte consumption < 4 pg) within 21 min. Thus, a few potential inhibitors of S-protein-ACE2 binding were identified. This includes (2S,3aS,6aS)-1-((S)-N-((S)-1-Carboxy-3-phenylpropyl)alanyl)tetrahydrocyclopenta[b] pyrrole-2-carboxylic acid (ramiprilat) and (2S,3aS,7aS)-1-[(2S)-2-[[(2S)-1-Carboxybutyl]amino]propanoyl]-2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (perindoprilat) that reduced the binding affinity of S-protein to ACE2 by 72% and 67%; and SARS-CoV-2 in vitro infectivity to the ACE2-expressing human oral cavity squamous carcinoma cells (OEC-M1) by 36.4 and 20.1%, respectively, compared to the PBS control. These findings demonstrated the usefulness of the developed biosensing platform for the rapid screening of modulators for S-protein-ACE2 binding.
Keywords: ACE2-SARS CoV 2 S-Protein interaction; Biosensor; Electrochemical impedance spectroscopy (EIS); Palladium nano-thin-film electrode; SARS-CoV-2 infection inhibitors.
【저자키워드】 biosensor, electrochemical impedance spectroscopy (EIS), ACE2-SARS CoV 2 S-Protein interaction, Palladium nano-thin-film electrode, SARS-CoV-2 infection inhibitors., 【초록키워드】 COVID-19, coronavirus disease, severe acute respiratory syndrome coronavirus 2, SARS-CoV-2, Coronavirus disease 2019, ACE2, coronavirus, SARS-COV-2 infection, progression, in vitro, severe acute respiratory syndrome Coronavirus, angiotensin-converting enzyme 2, inhibitors, binding affinity, Spike protein, biosensor, CoV, inhibitor, platform, binding, Angiotensin-converting enzyme, S-protein, Concentration, angiotensin, ACE2 binding, oral cavity, Interference, Volume, acute respiratory syndrome, acute respiratory syndrome coronavirus, cell surface, SARS CoV, palladium, Cell, pyrrole, PYRROLE-2-CARBOXYLIC ACID, detect, include, reduced, demonstrated, PBS, the binding affinity, attenuate, SARS-CoV-2 cell, 【제목키워드】 SARS-CoV-2, inhibitor, development, flexible,