TY - JOUR
T1 - SARS-CoV-2 Nucleocapsid Protein TR-FRET Assay Amenable to High Throughput Screening
AU - Gorshkov, Kirill
AU - Morales Vasquez, Desarey
AU - Chiem, Kevin
AU - Ye, Chengjin
AU - Nguyen Tran, Bruce
AU - Carlos De La Torre, Juan
AU - Moran, Thomas
AU - Chen, Catherine Z.
AU - Martinez-Sobrido, Luis
AU - Zheng, Wei
N1 - Publisher Copyright:
©
PY - 2022/1/14
Y1 - 2022/1/14
N2 - 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 (IC50) 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.
AB - 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 (IC50) 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.
KW - SARS-CoV-2
KW - TR-FRET
KW - assay development
KW - nucleocapsid phosphoprotein
KW - variants of concern
UR - http://www.scopus.com/inward/record.url?scp=85122734695&partnerID=8YFLogxK
U2 - 10.1021/acsptsci.1c00182
DO - 10.1021/acsptsci.1c00182
M3 - Article
AN - SCOPUS:85122734695
SN - 2575-9108
VL - 5
SP - 8
EP - 19
JO - ACS Pharmacology and Translational Science
JF - ACS Pharmacology and Translational Science
IS - 1
ER -