title: Development of a homogeneous high-throughput screening assay for biological inhibitors of human rhinovirus infection.
abstract: Infection with human rhinovirus (HRV) is thought to result in acute respiratory exacerbations of chronic obstructive pulmonary disorder (COPD). Consequently, prevention of HRV infection may provide therapeutic benefit to these patients. As all major group HRV serotypes infect cells via an interaction between viral coat proteins and intercellular adhesion molecule-1 (ICAM-1), it is likely that inhibitors of this interaction would prevent or reduce infections. Our objective was to use phage display technology in conjunction with naive human antibody libraries to identify anti-ICAM-1 antibodies capable of functional blockade of HRV infection. Key to success was the development of a robust, functionally relevant high-throughput screen (HTS) compatible with the specific challenges of antibody screening. In this article, we describe the development of a novel homogeneous time-resolved fluorescence (HTRF) assay based on the inhibition of soluble ICAM-1 binding to live HRV16. We describe the implementation of the method in an antibody screening campaign and demonstrate the biological relevance of the assay by confirming the activity of resultant antibodies in a cell-based in vitro HRV infection assay.
authors.affiliation: MedImmune Ltd., Cambridge CB21 6GH, UK. newtonp@medimmune.com
authors.forename: Philip
authors.initials: P
authors.lastname: Newton
authors.forename: Desmond
authors.initials: D
authors.lastname: O'Shea
authors.forename: Edward
authors.initials: E
authors.lastname: Wells
authors.forename: Kerry
authors.initials: K
authors.lastname: Moakes
authors.forename: Rebecca
authors.initials: R
authors.lastname: Dunmore
authors.forename: Robin J
authors.initials: RJ
authors.lastname: Butler
authors.forename: Trevor
authors.initials: T
authors.lastname: Wilkinson
authors.forename: Alison
authors.initials: A
authors.lastname: Ward
authors.forename: Nigel
authors.initials: N
authors.lastname: Casson
authors.forename: Martin
authors.initials: M
authors.lastname: Strain
authors.forename: Katherine
authors.initials: K
authors.lastname: Vousden
authors.forename: David C
authors.initials: DC
authors.lastname: Lowe
authors.forename: Debbie V
authors.initials: DV
authors.lastname: Pattison
authors.forename: Alan M
authors.initials: AM
authors.lastname: Carruthers
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Sleeman
authors.forename: Tristan J
authors.initials: TJ
authors.lastname: Vaughan
authors.forename: Paula
authors.initials: P
authors.lastname: Harrison
mesh_terms: D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D005453:Fluorescence; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism; D010850:Picornaviridae Infections / Q000276:immunology* / Q000378:metabolism; D012229:Rhinovirus / Q000276:immunology* / Q000378:metabolism
chemical_list: D000906:Antibodies; D018799:Intercellular Adhesion Molecule-1