title: FAITH - Fast Assembly Inhibitor Test for HIV.
abstract: Due to the high number of drug-resistant HIV-1 mutants generated by highly active antiretroviral therapy (HAART), there is continuing demand for new types of inhibitors. Both the assembly of the Gag polyprotein into immature and mature HIV-1 particles are attractive candidates for the blocking of the retroviral life cycle. Currently, no therapeutically-used assembly inhibitor is available. One possible explanation is the lack of a reliable and simple assembly inhibitor screening method. To identify compounds potentially inhibiting the formation of both types of HIV-1 particles, we developed a new fluorescent high-throughput screening assay. This assay is based on the quantification of the assembly efficiency in vitro in a 96-well plate format. The key components of the assay are HIV-1 Gag-derived proteins and a dual-labelled oligonucleotide, which emits fluorescence only when the assembly of retroviral particles is inhibited. The method was validated using three (CAI, BM2, PF74) reported assembly inhibitors.
authors.affiliation: Institute of Organic Chemistry and Biochemistry IOCB Research Centre & Gilead Sciences, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Prague, Czech Republic.
authors.forename: Romana
authors.initials: R
authors.lastname: Hadravová
authors.affiliation: Institute of Organic Chemistry and Biochemistry IOCB Research Centre & Gilead Sciences, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Prague, Czech Republic; Department of Biotechnology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague, Czech Republic. Electronic address: michaela.rumlova@vscht.cz.
authors.forename: Michaela
authors.initials: M
authors.lastname: Rumlová
authors.affiliation: Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 3, 166 28 Prague, Czech Republic. Electronic address: tomas.ruml@vscht.cz.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Ruml
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015683:Gene Products, gag / Q000378:metabolism; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D019065:Virus Assembly / Q000187:drug effects*
chemical_list: D019380:Anti-HIV Agents; D015683:Gene Products, gag
keywords: Assay; Assembly; CA; Capsid; HIV; High-throughput screening; Inhibitor; Retrovirus