title: Preexisting Antibodies to an F(ab')2 Antibody Therapeutic and Novel Method for Immunogenicity Assessment.
abstract: Anti-therapeutic antibodies (ATAs) may impact drug exposure and activity and induce immune complex mediated toxicity; therefore the accurate measurement of ATA is important for the analysis of drug safety and efficacy. Preexisting ATAs to the hinge region of anti-Delta like ligand 4 (anti-DLL4) F(ab')2, a potential antitumor therapeutic, were detected in cynomolgus monkey serum, which presented a challenge in developing assays for detecting treatment induced ATA. A total ATA assay was developed using a bridging ELISA that detected both anti-CDR and anti-framework ATA including anti-hinge reactivity. A competition assay that could detect 500 ng/mL of anti-CDR ATA in the presence of preexisting ATA was also developed to determine ATA specific to the anti-DLL4 F(ab')2 CDR using anti-DLL4 F(ab')2 and a control F(ab')2. We used these assay methods in a cynomolgus monkey in vivo study to successfully evaluate total and anti-CDR ATA. The preexisting anti-hinge reactivity was also observed to a lesser extent in human serum, and a similar approach could be applied for specific immunogenicity assessment in clinical trials.
authors.affiliation: BioAnalytical Sciences Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Jane
authors.identifier: 0000-0001-8839-1306
authors.initials: J
authors.lastname: Ruppel
authors.affiliation: BioAnalytical Sciences Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Ann
authors.initials: A
authors.lastname: Brady
authors.affiliation: BioAnalytical Sciences Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Rebecca
authors.identifier: 0000-0002-4372-1815
authors.initials: R
authors.lastname: Elliott
authors.affiliation: BioAnalytical Sciences Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Leddy
authors.affiliation: Biosample and Repository Management Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Marco
authors.identifier: 0000-0003-4157-6463
authors.initials: M
authors.lastname: Palencia
authors.affiliation: Nonclinical Biostatistics Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Coleman
authors.affiliation: Safety Assessment Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Couch
authors.affiliation: BioAnalytical Sciences Department, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors.forename: Eric
authors.identifier: 0000-0003-1793-9076
authors.initials: E
authors.lastname: Wakshull
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000009:adverse effects / Q000276:immunology* / Q000627:therapeutic use; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D007518:Isoantibodies / Q000097:blood / Q000276:immunology*; D008297:Male; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007518:Isoantibodies