Journal article
Development of antibody-based assays for high throughput discovery and mechanistic study of antiviral agents against yellow fever virus
Antiviral research, v 182, pp 104907-104907
01 Oct 2020
PMID: 32798604
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Despite the availability of a highly effective yellow fever virus (YFV) vaccine, outbreaks of yellow fever frequently occur in Africa and South America with significant mortality, highlighting the pressing need for antiviral drugs to manage future outbreaks. To support the discovery and development of antiviral drugs against YFV, we characterized a panel of rabbit polyclonal antibodies against the three YFV structural proteins and five non-structural proteins and demonstrated these antibody reagents in conjunction with viral RNA metabolic labeling, double-stranded RNA staining and membrane floatation assays as powerful tools for investigating YFV polyprotein processing, replication complex formation, viral RNA synthesis and high throughput discovery of antiviral drugs. Specifically, the proteolytic processing of the viral polyprotein can be analyzed by Western blot assays. The predominant nuclear localization of N55 protein as well as the relationship between intracellular viral non-structural protein distribution and foci of YFV RNA replication can be revealed by immunofluorescence staining and membrane flotation assays. Using an antibody against YFV NS4B protein as an example, in-cell western and high-content imaging assays have been developed for high throughput discovery of antiviral agents. A synergistic antiviral effect of an YFV NS4B-targeting antiviral agent BDAA and a N55 RNA-dependent RNA polymerase inhibitor (Sofosbuvir) was also demonstrated with the high-content imaging assay. Apparently, the antibody-based assays established herein not only facilitate the discovery and development of antiviral agents against YFV, but also provide valuable tools to dissect the molecular mechanism by which the antiviral agents inhibit YFV replication.
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Details
- Title
- Development of antibody-based assays for high throughput discovery and mechanistic study of antiviral agents against yellow fever virus
- Creators
- Zhao Gao - Baruch S. Blumberg InstituteLin Zhang - Baruch S. Blumberg InstituteJulia Ma - Baruch S. Blumberg InstituteAndrea Jurado - Rockefeller UniversitySeon-Hui Hong - Rockefeller UniversityJu-Tao Guo - Baruch S. Blumberg InstituteCharles M. Rice - Rockefeller UniversityMargaret R. MacDonald - Rockefeller UniversityJinhong Chang - Baruch S. Blumberg Institute
- Publication Details
- Antiviral research, v 182, pp 104907-104907
- Publisher
- Elsevier
- Number of pages
- 12
- Grant note
- Commonwealth of Pennsylvania through the Hepatitis B Foundation AI134732; AI124690 / National Institutes of Health, USA; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology
- Web of Science ID
- WOS:000576075400005
- Scopus ID
- 2-s2.0-85089751328
- Other Identifier
- 991020547321504721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Pharmacology & Pharmacy
- Virology