An innovative and highly drug-tolerant approach for detecting neutralizing antibodies directed to therapeutic antibodies.
OBJECTIVE
Immunogenicity testing of biotherapeutic drugs is a regulatory requirement. Herein, we describe a drug-tolerant assay for detecting neutralizing antibodies against a therapeutic antibody.


RESULTS
Excess target of the therapeutic antibody was incorporated into the detection step of an affinity capture elution assay. Signal generated from binding of antidrug antibody (ADA) to the therapeutic antibody was compared with signal from binding of ADA to the therapeutic antibody preincubated with its target. The results demonstrated that the target blocked binding of the therapeutic antibody to neutralizing monkey ADA and to two anti-idiotypic antibodies.


CONCLUSIONS
This highly drug-tolerant novel approach enables the detection of neutralizing antibodies and allows for one basic assay format to achieve complete characterization of ADA responses.
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, IN 46285, USA.
John H
JH
Sloan
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, IN 46285, USA.
Richard G
RG
Conway
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, IN 46285, USA.
Thomas G
TG
Pottanat
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, IN 46285, USA.
Jason S
JS
Troutt
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, IN 46285, USA.
Richard E
RE
Higgs
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, IN 46285, USA.
Robert J
RJ
Konrad
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, IN 46285, USA.
Yue-Wei
YW
Qian
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000097:blood* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000097:blood* / Q000276:immunology; D001710:Biotin / Q000737:chemistry; D002846:Chromatography, Affinity; D000882:Haplorhini; D006801:Humans; D007118:Immunoassay
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D001710:Biotin
affinity capture elution; drug tolerance; immunoassay; immunogenicity; neutralizing antidrug antibody; therapeutic antibody