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SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay Amenable to High Throughput Screening
[Image: see text] 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 been put forth, only a few compounds are in l...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8751018/ https://www.ncbi.nlm.nih.gov/pubmed/35036857 http://dx.doi.org/10.1021/acsptsci.1c00182 |
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author | Gorshkov, Kirill Morales Vasquez, Desarey Chiem, Kevin Ye, Chengjin Nguyen Tran, Bruce Carlos de la Torre, Juan Moran, Thomas Chen, Catherine Z. Martinez-Sobrido, Luis Zheng, Wei |
author_facet | Gorshkov, Kirill Morales Vasquez, Desarey Chiem, Kevin Ye, Chengjin Nguyen Tran, Bruce Carlos de la Torre, Juan Moran, Thomas Chen, Catherine Z. Martinez-Sobrido, Luis Zheng, Wei |
author_sort | Gorshkov, Kirill |
collection | PubMed |
description | [Image: see text] 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 been put forth, only a few compounds are 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 conjugated to donor and acceptor fluorophores that produce a robust ratiometric signal for high throughput screening of large compound collections. Using this assay, we measured a half maximal inhibitory concentration (IC(50)) for remdesivir of 9.3 μM against infection with SARS-CoV-2 USA/WA1/2020 (WA-1). The assay also detected SARS-CoV-2 South African (Beta, β), Brazilian/Japanese P.1 (Gamma, γ), and Californian (Epsilon, ε) variants of concern (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. |
format | Online Article Text |
id | pubmed-8751018 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-87510182022-01-11 SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay Amenable to High Throughput Screening Gorshkov, Kirill Morales Vasquez, Desarey Chiem, Kevin Ye, Chengjin Nguyen Tran, Bruce Carlos de la Torre, Juan Moran, Thomas Chen, Catherine Z. Martinez-Sobrido, Luis Zheng, Wei ACS Pharmacol Transl Sci [Image: see text] 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 been put forth, only a few compounds are 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 conjugated to donor and acceptor fluorophores that produce a robust ratiometric signal for high throughput screening of large compound collections. Using this assay, we measured a half maximal inhibitory concentration (IC(50)) for remdesivir of 9.3 μM against infection with SARS-CoV-2 USA/WA1/2020 (WA-1). The assay also detected SARS-CoV-2 South African (Beta, β), Brazilian/Japanese P.1 (Gamma, γ), and Californian (Epsilon, ε) variants of concern (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. American Chemical Society 2022-01-03 /pmc/articles/PMC8751018/ /pubmed/35036857 http://dx.doi.org/10.1021/acsptsci.1c00182 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Gorshkov, Kirill Morales Vasquez, Desarey Chiem, Kevin Ye, Chengjin Nguyen Tran, Bruce Carlos de la Torre, Juan Moran, Thomas Chen, Catherine Z. Martinez-Sobrido, Luis Zheng, Wei SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay Amenable to High Throughput Screening |
title | SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay
Amenable to High Throughput Screening |
title_full | SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay
Amenable to High Throughput Screening |
title_fullStr | SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay
Amenable to High Throughput Screening |
title_full_unstemmed | SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay
Amenable to High Throughput Screening |
title_short | SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay
Amenable to High Throughput Screening |
title_sort | sars-cov-2 nucleocapsid protein tr-fret assay
amenable to high throughput screening |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8751018/ https://www.ncbi.nlm.nih.gov/pubmed/35036857 http://dx.doi.org/10.1021/acsptsci.1c00182 |
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