A novel HIV-1 antiviral high throughput screening approach for the discovery of HIV-1 inhibitors.
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.
Blair
Wade S
WS
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
Isaacson
Jason
J
Li
Xinqiang
X
Cao
Joan
J
Peng
Qinghai
Q
Kong
George F Z
GF
Patick
Amy K
AK
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
D019380:Anti-HIV Agents; D049409:Luciferases, Firefly