A nanoluciferase SARS-CoV-2 for rapid neutralization testing and screening of anti-infective drugs for COVID-19

Nat Commun. 2020 Oct 15;11(1):5214. doi: 10.1038/s41467-020-19055-7.


A high-throughput platform would greatly facilitate coronavirus disease 2019 (COVID-19) serological testing and antiviral screening. Here we present a high-throughput nanoluciferase severe respiratory syndrome coronavirus 2 (SARS-CoV-2-Nluc) that is genetically stable and replicates similarly to the wild-type virus in cell culture. SARS-CoV-2-Nluc can be used to measure neutralizing antibody activity in patient sera within 5 hours, and it produces results in concordance with a plaque reduction neutralization test (PRNT). Additionally, using SARS-CoV-2-Nluc infection of A549 cells expressing human ACE2 receptor (A549-hACE2), we show that the assay can be used for antiviral screening. Using the optimized SARS-CoV-2-Nluc assay, we evaluate a panel of antivirals and other anti-infective drugs, and we identify nelfinavir, rupintrivir, and cobicistat as the most selective inhibitors of SARS-CoV-2-Nluc (EC50 0.77 to 2.74 µM). In contrast, most of the clinically approved antivirals, including tenofovir alafenamide, emtricitabine, sofosbuvir, ledipasvir, and velpatasvir were inactive at concentrations up to 10 µM. Collectively, this high-throughput platform represents a reliable tool for rapid neutralization testing and antiviral screening for SARS-CoV-2.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Angiotensin-Converting Enzyme 2
  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Viral / blood
  • Antiviral Agents / pharmacology*
  • Betacoronavirus / drug effects*
  • Betacoronavirus / genetics
  • Betacoronavirus / immunology
  • Betacoronavirus / isolation & purification*
  • COVID-19
  • Chlorocebus aethiops
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / virology
  • High-Throughput Screening Assays / methods*
  • Humans
  • Luciferases / genetics
  • Neutralization Tests / methods*
  • Pandemics
  • Peptidyl-Dipeptidase A / metabolism
  • Pneumonia, Viral / diagnosis*
  • Pneumonia, Viral / virology
  • SARS-CoV-2
  • Vero Cells
  • Virus Internalization / drug effects
  • Virus Replication / drug effects


  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Antiviral Agents
  • Luciferases
  • Peptidyl-Dipeptidase A
  • ACE2 protein, human
  • Angiotensin-Converting Enzyme 2