title: Development of a human papillomavirus competitive luminex immunoassay for 9 HPV types.
abstract: In the clinical trials of the quadrivalent human papillomavirus (qHPV) vaccine, antibodies were measured by a competitive Luminex immunoassay (HPV-4 cLIA). A nine-valent HPV (9vHPV) vaccine targeting the types in the qHPV vaccine (HPV6/11/16/18), as well as 5 of the next most frequent HPV types found in cervical cancers worldwide (HPV31/33/45/52/58) is under development. To support the 9vHPV vaccine program, a nine-multiplexed cLIA (HPV-9 cLIA) was developed. Antibody titers were determined in a competitive format, where type-specific phycoerythrin (PE)-labeled, neutralizing mAbs (mAbs-PE) compete with an individual's serum antibodies for binding to conformationally sensitive, neutralizing epitopes on the VLPs. Neutralizing antibody levels were quantitated against a reference standard - a pool of sera from 6 Rhesus macaques that were immunized with the 9vHPV vaccine. Specificity of the mAbs was assessed by measuring their individual binding capacities to the type-specific and non-type-specific VLPs at alternative concentrations of the mAbs. Antibody assignments to the HPV-9 cLIA reference standard for HPV6/11/16/18 were determined to provide for a measure of consistency in serostatus assignment between the HPV-4 and HPV-9 cLIAs. Antibody assignments to the HPV-9 reference standard for HPV31/33/45/52/58 were obtained by calibration to HPV11 using a direct binding IgG assay. For each HPV VLP type, the cross-reactivity of the mAb-PEs in the HPV-9 cLIA was <1% (i.e., the mAb-PEs result in <1% non-specific binding). The antibody concentrations assigned to the HPV-9 cLIA reference standard for types 6/11/16/18/31/33/45/52/58 were 3,817, 2,889, 23,061, 5,271, 3,942, 2,672, 1,489, 1274, and 2263 mMU/mL, respectively.
authors.affiliation: a Merck & Co., Inc.; Whitehouse Station, NJ USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Roberts
authors.forename: Tina
authors.initials: T
authors.lastname: Green
authors.forename: Erica
authors.initials: E
authors.lastname: Hess
authors.forename: Katie
authors.initials: K
authors.lastname: Matys
authors.forename: Martha J
authors.initials: MJ
authors.lastname: Brown
authors.forename: Richard M
authors.initials: RM
authors.lastname: Haupt
authors.forename: Alain
authors.initials: A
authors.lastname: Luxembourg
authors.forename: Scott
authors.initials: S
authors.lastname: Vuocolo
authors.forename: Alfred
authors.initials: A
authors.lastname: Saah
authors.forename: Joseph
authors.initials: J
authors.lastname: Antonello
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003429:Cross Reactions; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000097:blood; D008253:Macaca mulatta; D027383:Papillomaviridae / Q000276:immunology*; D030361:Papillomavirus Infections / Q000276:immunology* / Q000821:virology*; D012680:Sensitivity and Specificity; D022701:Virosomes
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D022701:Virosomes
keywords: Luminex; assay; human papillomavirus; vaccine