title: Refined analysis of antigen-specific antibody responses--a new one-step tool in immunogenicity studies.
abstract: Immunogenicity is a continuous efficacy and safety issue of biopharmaceuticals. Pre-clinical models for prediction of immunogenicity itself as well as biomarkers to reveal potential mechanisms behind an already existing antibody response are still needed. A sensitive, robust and specific immunogenicity assay has therefore been developed that can detect and measure antibodies of five classes against an administered recombinant human protein drug. Additionally, a validation was performed to evaluate the reproducibility and specificity of this newly developed assay. The production of drug-induced antibodies in mice injected with a recombinant human protein drug has been measured by using a modified version of a multi-parametric bead analysis technique. Competitive binding was used to verify drug-specificity of the antibodies. Results showed that the mouse response against the recombinant human protein was IgG1- and IgG2b-specific, suggesting that the drug-induced response was driven by both Th1/Th2 cells; a finding confirmed by measurement of the cytokine profile. With this assay, anti-drug antibody class and subclass screening may be executed in one step.
authors.affiliation: Immunotoxicology Group, Department of Molecular Toxicology, AstraZeneca, SE-151 85 Södertälje, Sweden. britta.granath@astrazeneca.com
authors.forename: Britta
authors.initials: B
authors.lastname: Granath
authors.forename: Jan
authors.initials: J
authors.lastname: Holgersson
authors.forename: Nina
authors.initials: N
authors.lastname: Brenden
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology*; D001702:Biopharmaceutics / Q000379:methods*; D019921:Biotinylation; D016207:Cytokines / Q000097:blood; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000379:methods; D015507:Drugs, Investigational / Q000009:adverse effects* / Q000737:chemistry / Q000592:standards; D005260:Female; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology
chemical_list: D000941:Antigens; D016207:Cytokines; D015507:Drugs, Investigational; D007074:Immunoglobulin G