Development and validation of a high-throughput screen for inhibitors of SARS CoV and its application in screening of a 100,000-compound library

William E. Severson, Nice Shindo, Mindy Sosa, Thomas Fletcher, E. Lucile White, Subramaniam Ananthan, Colleen Jonsson

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The authors have developed a high-throughput screen (HTS) that allows for the identification of potential inhibitors of the severe acute respiratory syndrome coronavirus (SARS CoV) from large compound libraries. The luminescent-based assay measures the inhibition of SARS CoV-induced cytopathic effect (CPE) in Vero E6 cells. The assay was validated in 96-well plates in a BSL3 containment facility. The assay is sensitive and robust, with Z values > 0.6, signal to background (S/B) > 16, and signal to noise (S/N) > 3. The assay was further validated with 2 different diversity sets of compounds against the SARS CoV. The "hit" rate for both libraries was approximately 0.01%. The validated HTS assay was then employed to screen a 100,000-compound library against SARS CoV. The hit rate for the library in a single-dose format was determined to be approximately 0.8%. Screening of the 3 libraries resulted in the identification of several novel compounds that effectively inhibited the CPE of SARS CoV in vitro - compounds which will serve as excellent lead candidates for further evaluation. At a 10-μM concentration, 3 compounds with selective indexes (SI50) of > 53 were discovered.

Original languageEnglish (US)
Pages (from-to)33-40
Number of pages8
JournalJournal of Biomolecular Screening
Volume12
Issue number1
DOIs
StatePublished - Feb 1 2007

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Severe Acute Respiratory Syndrome
Coronavirus
Libraries
Assays
Screening
Throughput
SARS Virus
Luminescent Measurements
Vero Cells
Noise

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

Cite this

Development and validation of a high-throughput screen for inhibitors of SARS CoV and its application in screening of a 100,000-compound library. / Severson, William E.; Shindo, Nice; Sosa, Mindy; Fletcher, Thomas; White, E. Lucile; Ananthan, Subramaniam; Jonsson, Colleen.

In: Journal of Biomolecular Screening, Vol. 12, No. 1, 01.02.2007, p. 33-40.

Research output: Contribution to journalArticle

Severson, William E. ; Shindo, Nice ; Sosa, Mindy ; Fletcher, Thomas ; White, E. Lucile ; Ananthan, Subramaniam ; Jonsson, Colleen. / Development and validation of a high-throughput screen for inhibitors of SARS CoV and its application in screening of a 100,000-compound library. In: Journal of Biomolecular Screening. 2007 ; Vol. 12, No. 1. pp. 33-40.
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