title: Development of a competitive enzyme linked immunosorbent assay to identify epitope specific antibodies in recipients of the U.S. licensed anthrax vaccine.
abstract: Vaccination with anthrax vaccine adsorbed (AVA) results in the production of protective antigen (PA) specific antibodies, which play an important protective role against anthrax toxins. Analyzing the specificity of serum antibodies generated in response to AVA vaccination can provide insight into the mechanisms of protective immunity against this important pathogen. The goal of this study was to develop a competitive enzyme linked immunosorbent assay (cELISA) to test human immune serum for antibodies specific for a known lethal toxin neutralizing epitope in PA. PA-specific antibodies in sera from individuals who received the six-dose AVA vaccine series competed for binding to immobilized PA with monoclonal antibody F20G75, which binds to a linear epitope in domain 2 of PA and neutralizes lethal toxin activity in vitro. These results suggest that antibodies in human AVA vaccinee serum recognize the same epitope as F20G75, or one in close proximity to it, and may serve a protective role against anthrax lethal toxin. This assay may be used for serological confirmation of successful immunization against anthrax and for the identification of antibodies in human vaccinee serum that recognize protective epitopes on PA.
authors.affiliation: Division of Vaccine Preventable Bacterial Diseases, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB, Canada. mike_gubbins@phac-aspc.gc.ca
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Gubbins
authors.forename: Lisa
authors.initials: L
authors.lastname: Schmidt
authors.forename: Raymond S
authors.initials: RS
authors.lastname: Tsang
authors.forename: Jody D
authors.initials: JD
authors.lastname: Berry
authors.forename: Amin
authors.initials: A
authors.lastname: Kabani
authors.forename: Donald I H
authors.initials: DI
authors.lastname: Stewart
mesh_terms: D000595:Amino Acid Sequence; D022122:Anthrax Vaccines / Q000008:administration & dosage / Q000276:immunology*; D000907:Antibodies, Bacterial / Q000097:blood*; D000911:Antibodies, Monoclonal / Q000235:genetics; D000918:Antibody Specificity / Q000276:immunology*; D000942:Antigens, Bacterial / Q000276:immunology; D001427:Bacterial Toxins / Q000276:immunology; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018604:Epitope Mapping / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics; D014481:United States
chemical_list: D022122:Anthrax Vaccines; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D010455:Peptides; C030325:anthrax toxin