tetano
Editor, Senior Moderator
ACS Pharmacol Transl Sci
. 2022 Jan 3;5(1):8-19.
doi: 10.1021/acsptsci.1c00182. eCollection 2022 Jan 14.
SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay Amenable to High Throughput Screening
Kirill Gorshkov[SUP] 1 [/SUP], Desarey Morales Vasquez[SUP] 2 [/SUP], Kevin Chiem[SUP] 2 [/SUP], Chengjin Ye[SUP] 2 [/SUP], Bruce Nguyen Tran[SUP] 1 [/SUP], Juan Carlos de la Torre[SUP] 3 [/SUP], Thomas Moran[SUP] 4 [/SUP], Catherine Z Chen[SUP] 1 [/SUP], Luis Martinez-Sobrido[SUP] 2 [/SUP], Wei Zheng[SUP] 1 [/SUP]
Affiliations
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 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[SUB]50[/SUB]) 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.
. 2022 Jan 3;5(1):8-19.
doi: 10.1021/acsptsci.1c00182. eCollection 2022 Jan 14.
SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay Amenable to High Throughput Screening
Kirill Gorshkov[SUP] 1 [/SUP], Desarey Morales Vasquez[SUP] 2 [/SUP], Kevin Chiem[SUP] 2 [/SUP], Chengjin Ye[SUP] 2 [/SUP], Bruce Nguyen Tran[SUP] 1 [/SUP], Juan Carlos de la Torre[SUP] 3 [/SUP], Thomas Moran[SUP] 4 [/SUP], Catherine Z Chen[SUP] 1 [/SUP], Luis Martinez-Sobrido[SUP] 2 [/SUP], Wei Zheng[SUP] 1 [/SUP]
Affiliations
- PMID: 35036857
- PMCID: PMC8751018
- DOI: 10.1021/acsptsci.1c00182
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 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[SUB]50[/SUB]) 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.