Improving anti-drug antibody assay performance in Gyrolab for therapeutic recombinant antibody Infliximab

Abstract: Monoclonal antibodies can be used as targeting therapies for several diseases. One major concern when using these therapies is anti-drug antibodies which may hamper the drugs efficiency. Gyrolab is an automated platform which can be used to develop bridging immunoassays where the anti-drug antibodies affinity towards the monoclonal antibody is utilized. Anti-drug antibody immunoassay development on Gyrolab is limited mainly by three factors which may inappropriately affect signal intensity levels. In this project different variants of bridging immunoassays based on drug Fab fragments have been developed for monoclonal antibody Infliximab, with the purpose to illustrate the effects of these three factors. Findings indicate that an assay based completely on drug Fab fragments is more sensitive compared to an assay based on intact drug since less affected by unspecific interactions between drug reagents and complex formations. Surprisingly findings also indicate that an assay based completely on drug Fab fragments is affected by human anti-hinge antibodies which decrease assay sensitivity. The most optimal assay variant is based on the combination between intact capture drug and Fab fragment as detection. This variant is insensitive to false positive reactions caused by Rheumatoid factor and human anti-hinge antibodies, less prone to form unspecific interactions between drug reagents and complex formations in the presence of anti-drug antibodies. The optimal assay variant also demonstrates best drug tolerance in combination with acid dissociation.