title: Development of a homogeneous screening assay for automated detection of antiviral agents active against severe acute respiratory syndrome-associated coronavirus.
abstract: The severity and global spread of the 2003 outbreak of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) highlighted the risks to human health posed by emerging viral diseases and emphasized the need for specific therapeutic agents instead of relying on existing broadly active antiviral compounds. The development of rapid screening assays is essential for antiviral drug discovery. Thus, a screening system for anti-SARS-CoV agents was developed, which evaluated compound potency, specificity and cytotoxicity at the initial screening phase. Cell lines were engineered to constitutively express an enhanced green fluorescent protein (EGFP) and used to detect (1) antiviral potency in SARS-CoV infection tests; (2) antiviral specificity in tests using the porcine coronavirus transmissible gastroenteritis virus (TGEV); and (3) cytotoxicity in the same assays without virus challenge. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. The suitability of this assay system in drug discovery was demonstrated by screening of 3388 small molecule compounds. The results show that these assays can be applied to high-throughput screening for identification of inhibitors selectively active against SARS-CoV.
authors.affiliation: Tibotec BVBA, Lead Discovery Operations, Gen De Wittelaan L 11B 3, Mechelen 2800, Belgium.
authors.forename: Tania
authors.initials: T
authors.lastname: Ivens
authors.forename: Christel
authors.initials: C
authors.lastname: Van den Eynde
authors.forename: Koen
authors.initials: K
authors.lastname: Van Acker
authors.forename: Erik
authors.initials: E
authors.lastname: Nijs
authors.forename: Géry
authors.initials: G
authors.lastname: Dams
authors.forename: Eva
authors.initials: E
authors.lastname: Bettens
authors.forename: Asa
authors.initials: A
authors.lastname: Ohagen
authors.forename: Rudi
authors.initials: R
authors.lastname: Pauwels
authors.forename: Kurt
authors.initials: K
authors.lastname: Hertogs
mesh_terms: D000998:Antiviral Agents / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D049452:Green Fluorescent Proteins; D045473:Severe acute respiratory syndrome-related coronavirus / Q000187:drug effects* / Q000502:physiology; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D049452:Green Fluorescent Proteins