title: Impact of matrix-associated soluble factors on the specificity of the immunogenicity assessment.
abstract: BACKGROUND
  Specificity and sensitivity are essential in assays for immunogenicity assessment of biotherapeutics. Nonspecific interactions from excess therapeutic or anti-therapeutic antibody, soluble ligands (e.g., target receptor), or serum proteins associated with autoimmune conditions (e.g., rheumatoid factor) in samples can impact the detection of a true anti-therapeutic response.
  
  
  RESULTS
  Electrochemiluminescence-based bridging assay formats could eliminate the interference due to rheumatoid factor with no pretreatment with Melon Gel™ or aggregated IgG. The interference due to soluble factors was not platform specific for the four therapeutics evaluated in this study.
  
  
  CONCLUSIONS
  Melon Gel pretreatment and avidin high-bind (Meso Scale Discovery) plates can effectively reduce interference due to rheumatoid factor in ELISA- and electrochemiluminescence-based assays, respectively. Excess levels of therapeutic and anti-therapeutic antibodies in bridging assays can impact assay specificity.
authors.affiliation: Clinical Immunology Department, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA.
authors.forename: Ami C
authors.initials: AC
authors.lastname: Bautista
authors.forename: Danika
authors.initials: D
authors.lastname: Wullner
authors.forename: Mike
authors.initials: M
authors.lastname: Moxness
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Swanson
authors.forename: Narendra
authors.initials: N
authors.lastname: Chirmule
authors.forename: Vibha
authors.initials: V
authors.lastname: Jawa
mesh_terms: D053000:Analytic Sample Preparation Methods; D000906:Antibodies / Q000097:blood / Q000276:immunology; D016477:Artifacts; D001681:Biological Assay / Q000379:methods*; D006801:Humans; D007109:Immunity / Q000187:drug effects*; D007118:Immunoassay; D008024:Ligands; D004364:Pharmaceutical Preparations / Q000032:analysis* / Q000378:metabolism; D012995:Solubility
chemical_list: D000906:Antibodies; D008024:Ligands; D004364:Pharmaceutical Preparations