title: A novel HIV-1 antiviral high throughput screening approach for the discovery of HIV-1 inhibitors.
abstract: Antiviral high throughput screens remain a viable option for identifying novel target inhibitors. However, few antiviral screens have been reduced to practice on an industrial scale. In this study, we describe an HIV-1 dual reporter assay that allows for the simultaneous evaluation of the potential antiviral activities and cytotoxicities of compounds in a high throughput screen (HTS) format. We validate the assay with known HIV-1 inhibitors and show that the antiviral and cytotoxic activities of compounds are reproducibly measured under screening conditions. In addition, we show that the assay exhibits parameters (e.g., signal-to-background ratios and Z' coefficients) suitable for high throughout screening. In a pilot screen, we demonstrate that non-specific or cytotoxic compounds represent a significant fraction of the hits identified in an antiviral screen and that these false positives are identified and deprioritized by the HIV-1 dual reporter assay at the primary screening step. We propose that the HIV-1 dual reporter assay represents a novel approach to HIV-1 antiviral screening that allows for the effective execution of industrial scale HTS campaigns with significantly greater returns on resource investment when compared to previous methods.
authors.affiliation: Department of Virology, Pfizer Global Research and Development, La Jolla Laboratories, 10777 Science Center Dr. (CB1), San Diego, CA 92121, USA. wade.blair@pfizer.com
authors.forename: Wade S
authors.initials: WS
authors.lastname: Blair
authors.forename: Jason
authors.initials: J
authors.lastname: Isaacson
authors.forename: Xinqiang
authors.initials: X
authors.lastname: Li
authors.forename: Joan
authors.initials: J
authors.lastname: Cao
authors.forename: Qinghai
authors.initials: Q
authors.lastname: Peng
authors.forename: George F Z
authors.initials: GF
authors.lastname: Kong
authors.forename: Amy K
authors.initials: AK
authors.lastname: Patick
mesh_terms: D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D002460:Cell Line; D017930:Genes, Reporter; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D049409:Luciferases, Firefly / Q000235:genetics; D008826:Microbial Sensitivity Tests / Q000379:methods* / Q000706:statistics & numerical data; D015203:Reproducibility of Results
chemical_list: D019380:Anti-HIV Agents; D049409:Luciferases, Firefly