title: Microarray compound screening (microARCS) to identify inhibitors of HIV integrase.
abstract: A novel high-throughput strand transfer assay has been developed, using Microarray Compound Screening (microARCS) technology, to identify inhibitors of human immunodeficiency virus (HIV) integrase. This technology utilizes agarose matrices to introduce a majority of the reagents throughout the assay. Integration of biotinylated donor DNA with fluorescein isothiocyanate (FITC)-labeled target DNA occurs on a SAM membrane in the presence of integrase. An anti-FITC antibody conjugated to alkaline phosphatase (AP) was used to do an enzyme-linked immunosorbent assay with the SAM. An agarose gel containing AttoPhos, a substrate of AP, was used for detection of the integrase reactions on the SAM. For detection, the AttoPhos gel was separated from the SAM after incubation and then the gel was imaged using an Eagle Eye II closed-circuit device camera system. Potential integrase inhibitors appear as dark spots on the gel image. A library of approximately 250,000 compounds was screened using this HIV integrase strand transfer assay in microARCS format. Compounds from different structural classes were identified in this assay as novel integrase inhibitors.
authors.affiliation: Department of Biological Screening, Abbott Laboratories, Global Pharmaceutical Products Division, Abbott Park, IL 60064, USA. Caroline.A.David@abbott.com
authors.forename: Caroline A
authors.initials: CA
authors.lastname: David
authors.forename: Tim
authors.initials: T
authors.lastname: Middleton
authors.forename: Debra
authors.initials: D
authors.lastname: Montgomery
authors.forename: Hock Ben
authors.initials: HB
authors.lastname: Lim
authors.forename: Warren
authors.initials: W
authors.lastname: Kati
authors.forename: Akhter
authors.initials: A
authors.lastname: Molla
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Xuei
authors.forename: Usha
authors.initials: U
authors.lastname: Warrior
authors.forename: James L
authors.initials: JL
authors.lastname: Kofron
authors.forename: David J
authors.initials: DJ
authors.lastname: Burns
mesh_terms: D019427:HIV Integrase / Q000187:drug effects*; D019428:HIV Integrase Inhibitors / Q000032:analysis* / Q000494:pharmacology
chemical_list: D019428:HIV Integrase Inhibitors; D019427:HIV Integrase