A cell-based luminescence assay is effective for high-throughput screening of potential influenza antivirals

James W. Noah, William Severson, Diana L. Noah, Lynn Rasmussen, E. Lucile White, Colleen Jonsson

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

The spread of highly pathogenic avian influenza across geographical and species barriers underscores the increasing need for novel antivirals to compliment vaccination and existing antiviral therapies. Identification of new antiviral lead compounds depends on robust primary assays for high-throughput screening (HTS) of large compound libraries. We have developed a cell-based screen for potential influenza antivirals that measures the cytopathic effect (CPE) induced by influenza virus (A/Udorn/72, H3N2) infection in Madin Darby canine kidney (MDCK) cells using the luminescent-based CellTiter Glo system. This 72 h assay is validated for HTS in 384-well plates and performs more consistently and reliably than methods using neutral red, with Z values > 0.8, signal-to-background > 30 and signal-to-noise > 10. In a blinded pilot screen (n = 10,781) at 10 μM concentration, four compounds (with previously demonstrated efficacy against influenza) inhibited viral-induced CPE by >50%, with EC50/CC50 values comparable to those determined by other cell-based assays, thereby validating this assay accuracy and ability to simultaneously evaluate compound cellular availability and/or toxicity. This assay is translatable for screening against other influenza strains, such as avian flu, and may facilitate identification of antivirals for other viruses that induce CPE, such as West Nile or Dengue.

Original languageEnglish (US)
Pages (from-to)50-59
Number of pages10
JournalAntiviral Research
Volume73
Issue number1
DOIs
StatePublished - Jan 1 2007

Fingerprint

Luminescence
Human Influenza
Antiviral Agents
Influenza in Birds
Viral Cytopathogenic Effect
High-Throughput Screening Assays
Neutral Red
Madin Darby Canine Kidney Cells
Dengue
Influenza A virus
Libraries
Noise
Vaccination
Viruses
Infection
Therapeutics

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Virology

Cite this

A cell-based luminescence assay is effective for high-throughput screening of potential influenza antivirals. / Noah, James W.; Severson, William; Noah, Diana L.; Rasmussen, Lynn; White, E. Lucile; Jonsson, Colleen.

In: Antiviral Research, Vol. 73, No. 1, 01.01.2007, p. 50-59.

Research output: Contribution to journalArticle

Noah, James W. ; Severson, William ; Noah, Diana L. ; Rasmussen, Lynn ; White, E. Lucile ; Jonsson, Colleen. / A cell-based luminescence assay is effective for high-throughput screening of potential influenza antivirals. In: Antiviral Research. 2007 ; Vol. 73, No. 1. pp. 50-59.
@article{be8027b5501b4bf1ac161564af280211,
title = "A cell-based luminescence assay is effective for high-throughput screening of potential influenza antivirals",
abstract = "The spread of highly pathogenic avian influenza across geographical and species barriers underscores the increasing need for novel antivirals to compliment vaccination and existing antiviral therapies. Identification of new antiviral lead compounds depends on robust primary assays for high-throughput screening (HTS) of large compound libraries. We have developed a cell-based screen for potential influenza antivirals that measures the cytopathic effect (CPE) induced by influenza virus (A/Udorn/72, H3N2) infection in Madin Darby canine kidney (MDCK) cells using the luminescent-based CellTiter Glo system. This 72 h assay is validated for HTS in 384-well plates and performs more consistently and reliably than methods using neutral red, with Z values > 0.8, signal-to-background > 30 and signal-to-noise > 10. In a blinded pilot screen (n = 10,781) at 10 μM concentration, four compounds (with previously demonstrated efficacy against influenza) inhibited viral-induced CPE by >50{\%}, with EC50/CC50 values comparable to those determined by other cell-based assays, thereby validating this assay accuracy and ability to simultaneously evaluate compound cellular availability and/or toxicity. This assay is translatable for screening against other influenza strains, such as avian flu, and may facilitate identification of antivirals for other viruses that induce CPE, such as West Nile or Dengue.",
author = "Noah, {James W.} and William Severson and Noah, {Diana L.} and Lynn Rasmussen and White, {E. Lucile} and Colleen Jonsson",
year = "2007",
month = "1",
day = "1",
doi = "10.1016/j.antiviral.2006.07.006",
language = "English (US)",
volume = "73",
pages = "50--59",
journal = "Antiviral Research",
issn = "0166-3542",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - A cell-based luminescence assay is effective for high-throughput screening of potential influenza antivirals

AU - Noah, James W.

AU - Severson, William

AU - Noah, Diana L.

AU - Rasmussen, Lynn

AU - White, E. Lucile

AU - Jonsson, Colleen

PY - 2007/1/1

Y1 - 2007/1/1

N2 - The spread of highly pathogenic avian influenza across geographical and species barriers underscores the increasing need for novel antivirals to compliment vaccination and existing antiviral therapies. Identification of new antiviral lead compounds depends on robust primary assays for high-throughput screening (HTS) of large compound libraries. We have developed a cell-based screen for potential influenza antivirals that measures the cytopathic effect (CPE) induced by influenza virus (A/Udorn/72, H3N2) infection in Madin Darby canine kidney (MDCK) cells using the luminescent-based CellTiter Glo system. This 72 h assay is validated for HTS in 384-well plates and performs more consistently and reliably than methods using neutral red, with Z values > 0.8, signal-to-background > 30 and signal-to-noise > 10. In a blinded pilot screen (n = 10,781) at 10 μM concentration, four compounds (with previously demonstrated efficacy against influenza) inhibited viral-induced CPE by >50%, with EC50/CC50 values comparable to those determined by other cell-based assays, thereby validating this assay accuracy and ability to simultaneously evaluate compound cellular availability and/or toxicity. This assay is translatable for screening against other influenza strains, such as avian flu, and may facilitate identification of antivirals for other viruses that induce CPE, such as West Nile or Dengue.

AB - The spread of highly pathogenic avian influenza across geographical and species barriers underscores the increasing need for novel antivirals to compliment vaccination and existing antiviral therapies. Identification of new antiviral lead compounds depends on robust primary assays for high-throughput screening (HTS) of large compound libraries. We have developed a cell-based screen for potential influenza antivirals that measures the cytopathic effect (CPE) induced by influenza virus (A/Udorn/72, H3N2) infection in Madin Darby canine kidney (MDCK) cells using the luminescent-based CellTiter Glo system. This 72 h assay is validated for HTS in 384-well plates and performs more consistently and reliably than methods using neutral red, with Z values > 0.8, signal-to-background > 30 and signal-to-noise > 10. In a blinded pilot screen (n = 10,781) at 10 μM concentration, four compounds (with previously demonstrated efficacy against influenza) inhibited viral-induced CPE by >50%, with EC50/CC50 values comparable to those determined by other cell-based assays, thereby validating this assay accuracy and ability to simultaneously evaluate compound cellular availability and/or toxicity. This assay is translatable for screening against other influenza strains, such as avian flu, and may facilitate identification of antivirals for other viruses that induce CPE, such as West Nile or Dengue.

UR - http://www.scopus.com/inward/record.url?scp=33846152578&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846152578&partnerID=8YFLogxK

U2 - 10.1016/j.antiviral.2006.07.006

DO - 10.1016/j.antiviral.2006.07.006

M3 - Article

VL - 73

SP - 50

EP - 59

JO - Antiviral Research

JF - Antiviral Research

SN - 0166-3542

IS - 1

ER -