title: Complex gel permeation assays for screening combinatorial libraries.
abstract: Gel permeation methods have been commonly used to screen combinatorial libraries synthesized on a solid support. We report here three screens of combinatorial libraries using gel permeation assays. These include a simple enzymatic assay to identify inhibitors of the influenza enzyme neuraminidase, and two more complex assays designed to screen for inhibitors of the interleukin-8 (IL-8)-IL-8 receptor and the urokinase-urokinase receptor interactions, respectively. The IL-8 ligand-receptor assay makes use of IL-8 receptor-expressing cells attached to a membrane, thus enabling washing steps as part of the assay. The urokinase ligand-receptor assay employs an enzyme-linked immunosorbent assay-type format, previously thought to be amenable only to well-based assays. The results of these three screens are reported here, including the discovery of a novel series of acyclic inhibitors of neuraminidase. The development of complex assays in a gel permeation format allows for the routine screening of combinatorially as well as noncombinatorially made compound collections against virtually any kind of target, and is being widely used in our high throughput screening operations.
authors.affiliation: Department of Biological Screening, Abbott Laboratories, Abbott Park, IL 60064-3500, USA. mark.schurdak@abbott.com
authors.forename: M E
authors.initials: ME
authors.lastname: Schurdak
authors.forename: M J
authors.initials: MJ
authors.lastname: Voorbach
authors.forename: L
authors.initials: L
authors.lastname: Gao
authors.forename: X
authors.initials: X
authors.lastname: Cheng
authors.forename: K M
authors.initials: KM
authors.lastname: Comess
authors.forename: S M
authors.initials: SM
authors.lastname: Rottinghaus
authors.forename: U
authors.initials: U
authors.lastname: Warrior
authors.forename: H N
authors.initials: HN
authors.lastname: Truong
authors.forename: D J
authors.initials: DJ
authors.lastname: Burns
authors.forename: B A
authors.initials: BA
authors.lastname: Beutel
mesh_terms: D002850:Chromatography, Gel / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D016209:Interleukin-8 / Q000378:metabolism; D009439:Neuraminidase / Q000037:antagonists & inhibitors; D009975:Orthomyxoviridae / Q000201:enzymology; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism; D023062:Receptors, Interleukin-8A / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D016209:Interleukin-8; D011956:Receptors, Cell Surface; D023062:Receptors, Interleukin-8A; D055293:Receptors, Urokinase Plasminogen Activator; D009439:Neuraminidase; D014568:Urokinase-Type Plasminogen Activator