A nanoluciferase SARS-CoV-2 for rapid neutralization testing and screening of anti-infective drugs for COVID-19
Science
Pneumonia, Viral
General Physics and Astronomy
Peptidyl-Dipeptidase A
Antibodies, Viral
Antiviral Agents
General Biochemistry, Genetics and Molecular Biology
Article
Betacoronavirus
Neutralization Tests
Chlorocebus aethiops
Animals
Humans
Luciferases
Pandemics
Vero Cells
SARS-CoV-2
Q
COVID-19
General Chemistry
Virus Internalization
Antibodies, Neutralizing
High-Throughput Screening Assays
3. Good health
A549 Cells
Angiotensin-Converting Enzyme 2
Coronavirus Infections
DOI:
10.1101/2020.06.22.165712
Publication Date:
2020-06-24T03:24:36Z
AUTHORS (15)
ABSTRACT
AbstractA high-throughput platform would greatly facilitate COVID-19 serological testing and antiviral screening. Here we report a nanoluciferase SARS-CoV-2 (SARS-CoV-2-Nluc) that is genetically stable and replicates similarly to the wild-type virus in cell culture. We demonstrate that the optimized reporter virus assay in Vero E6 cells can be used to measure neutralizing antibody activity in patient sera and produces results in concordance with a plaque reduction neutralization test (PRNT). Compared with the low-throughput PRNT (3 days), the SARS-CoV-2-Nluc assay has substantially shorter turnaround time (5 hours) with a high-throughput testing capacity. Thus, the assay can be readily deployed for large-scale vaccine evaluation and neutralizing antibody testing in humans. Additionally, we developed a high-throughput antiviral assay using SARS-CoV-2-Nluc infection of A549 cells expressing human ACE2 receptor (A549-hACE2). When tested against this reporter virus, remdesivir exhibited substantially more potent activity in A549-hACE2 cells compared to Vero E6 cells (EC500.115 vs 1.28 μM), while this difference was not observed for chloroquine (EC501.32 vs 3.52 μM), underscoring the importance of selecting appropriate cells for antiviral testing. Using the optimized SARS-CoV-2-Nluc assay, we evaluated a collection of approved and investigational antivirals and other anti-infective drugs. Nelfinavir, rupintrivir, and cobicistat were identified as the most selective inhibitors of SARS-CoV-2-Nluc (EC500.77 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.
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CITATIONS (15)
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