AIDSWEEKLY Plus; Monday, November 29, 2004
Staff Medical Writers

"This article describes the automation of an in vitro cell-based fusion assay for the identification of novel inhibitors of receptor mediated HIV-1 entry. The assay utilizes two stable cell lines: one expressing CD4, CCR5 and an LTR-promoter/beta-galactosidase reporter construct, and the other expressing gp160 and tat.

"Accumulation of beta-galactosidase can only occur following fusion of these two cell lines via the gp160 and receptor mediators, as this event facilitates the transfer of the tat transcription factor between the two cell types," scientists in England report.

"Although similar cell fusion systems have been described previously," wrote J. Bradley and coworkers, "they have not met the requirements for HTS due to complexity, throughput and reagent cost."

"The assay described in this article provides significant advantage, as (a) no transfection/infection events are required prior to the assay, reducing the potential for variability, (b) cells are mixed in solution, enhancing fusion efficiency compared to adherent cells, (c) miniaturization to low volume enables screening in 384-well plates; and (d) online cell dispensing facilitates automated screening," said the authors.

Bradley concluded, "This assay has been employed to screen approximately 650,000 compounds in a singleton format. The data demonstrate that the assay is robust, with a Z' consistently above 0.6, which compares favorably with less complex biochemical assays."

Bradley and colleagues published their study in the Journal of Biomolecular Screening (Development and automation of a 384-well cell fusion assay to identify inhibitors of CCR5/CD4-mediated HIV virus entry. J Biomol Screen. 2004 Sep;9(6):516-24.

For more information, contact C. Williams, Pfizer Ltd, Global Research and Development, HDG, IPC 654, Ramsgate Rd., Sandwich CT13 9NJ, Kent, England.

Publisher contact information for the Journal of Biomolecular Screening is: Sage Publications Inc., 2455 Teller Rd., Thousand Oaks, CA 91320, USA.

The information in this article comes under the major subject areas of HIV/AIDS, Pharmaceutical and Drug Development, Antiretroviral Therapy, and HIV Fusion Inhibitor.

This article was prepared by AIDS Weekly editors from staff and other reports.

Reference

Bradley J, Gill J, Bertelli F, et al., "Development and automation of a 384-well cell fusion assay to identify inhibitors of CCR5/CD4-mediated HIV virus entry", J Biomol Screen. 2004 Sep;9(6):516-24.

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