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A SARS-CoV-2 nucleocapsid protein TR-FRET assay amenable to high-throughput screening
Kirill Gorshkov; Desarey Morales Vasquez; Kevin Chiem; Chengjin Ye; Bruce Nguyen Tran; Juan Carlos de la Torre; Thomas Moran; Catherine Z. Chen; Luis Martinez-Sobrido; Wei Zheng.
Afiliación
  • Kirill Gorshkov; National Center for Advancing Translational Sciences
  • Desarey Morales Vasquez; Texas Biomedical Research Institute
  • Kevin Chiem; Texas Biomedical Research Institute
  • Chengjin Ye; Texas Biomedical Research Institute
  • Bruce Nguyen Tran; National Center for Advancing Translational Sciences
  • Juan Carlos de la Torre; Scripps Research Institute
  • Thomas Moran; Icahn School of Medicine at Mt. Sinai
  • Catherine Z. Chen; National Center for Advancing Translational Sciences
  • Luis Martinez-Sobrido; Texas Biomedical Research Institute
  • Wei Zheng; National Center for Advancing Translational Sciences
Preprint en En | PREPRINT-BIORXIV | ID: ppbiorxiv-450938
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ABSTRACT
Drug development for specific antiviral agents against coronavirus disease 2019 (COVID-19) is still an unmet medical need as the pandemic continues to spread globally. Although huge efforts for drug repurposing and compound screens have put forth, only few compounds remain in late stage clinical trials. New approaches and assays are needed to accelerate COVID-19 drug discovery and development. Here we report a time-resolved fluorescence resonance energy transfer-based assay that detects the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (NP) produced in infected cells. It uses two specific anti-NP monoclonal antibodies (MAbs) conjugated to donor and acceptor fluorophores that produces a robust ratiometric signal for high throughput screening of large compound collections. Using this assay, we measured a half maximal inhibitory concentration (IC50) for Remdesivir of 9.3 M against infection with SARS-CoV-2 USA/WA1/2020 (WA-1). The assay also detected SARS-CoV-2 South African (Beta, {beta}), Brazilian/Japanese variant P.1 (Gamma, {gamma}), and Californian (Epsilon, {varepsilon}), variants of concern or interest (VoC). Therefore, this homogeneous SARS-CoV-2 NP detection assay can be used for accelerating lead compound discovery for drug development and for evaluating drug efficacy against emerging SARS-CoV-2 VoC.
Licencia
cc_by_nc_nd
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Tipo de estudio: Diagnostic_studies / Experimental_studies / Prognostic_studies Idioma: En Año: 2021 Tipo del documento: Preprint
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Tipo de estudio: Diagnostic_studies / Experimental_studies / Prognostic_studies Idioma: En Año: 2021 Tipo del documento: Preprint