title: Identification and inhibition of drug target interference in immunogenicity assays.
abstract: A well-designed anti-drug antibody (ADA) immunoassay is critical for appropriately monitoring the immunogenicity profile of a therapeutic protein during its development. AMG 386 is a peptide-Fc fusion protein that inhibits angiogenesis by preventing the interaction of angiopoietins with the Tie2 receptor. In bridging immunoassays for ADA, interference by the drug target, present in the assay sample, can result in false positive antibody detection. We used a statistical design-of-experiments approach to identify angiopoietin interference in bridging immunoassays of anti-AMG 386 antibodies. We also demonstrated that a high-affinity monoclonal antibody, directed against an epitope on angiopoietin that competes with AMG 386 binding, could inhibit the angiopoietin interference while preserving the detection of ADA. This report describes the development and validation of methodologies for evaluating and addressing drug target interference in bioanalytical assays that involve interactions between drug, ADA, immune complexes, and drug target.
authors.affiliation: Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA. zhongz@amgen.com
authors.forename: Zhandong D
authors.initials: ZD
authors.lastname: Zhong
authors.forename: Steve
authors.initials: S
authors.lastname: Dinnogen
authors.forename: Martha
authors.initials: M
authors.lastname: Hokom
authors.forename: Chad
authors.initials: C
authors.lastname: Ray
authors.forename: David
authors.initials: D
authors.lastname: Weinreich
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Swanson
authors.forename: Narendra
authors.initials: N
authors.lastname: Chirmule
mesh_terms: D042682:Angiopoietins / Q000097:blood / Q000235:genetics / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D001681:Biological Assay; D002986:Clinical Trials as Topic; D004347:Drug Interactions; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007141:Immunoglobulin Fc Fragments / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D008297:Male; D051379:Mice; D009369:Neoplasms / Q000097:blood* / Q000188:drug therapy / Q000235:genetics / Q000276:immunology; D009389:Neovascularization, Pathologic / Q000097:blood* / Q000188:drug therapy / Q000276:immunology; D010455:Peptides / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D042787:Receptor, TIE-2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D042682:Angiopoietins; D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D010455:Peptides; D042787:Receptor, TIE-2