Emerging Technologies and Generic Assays for the Detection of Anti-Drug Antibodies

Abstract

Anti-drug antibodies induced by biologic therapeutics often impact drug pharmacokinetics, pharmacodynamics response, clinical efficacy, and patient safety. It is critical to assess the immunogenicity risk of potential biotherapeutics in producing neutralizing and nonneutralizing anti-drug antibodies, especially in clinical phases of drug development. Different assay methodologies have been used to detect all anti-drug antibodies, including ELISA, radioimmunoassay, surface plasmon resonance, and electrochemiluminescence-based technologies. The most commonly used method is a bridging assay, performed in an ELISA or on the Meso Scale Discovery platform. In this report, we aim to review the emerging new assay technologies that can complement or address challenges associated with the bridging assay format in screening and confirmation of ADAs. We also summarize generic anti-drug antibody assays that do not require drug-specific reagents for nonclinical studies. These generic assays significantly reduce assay development efforts and, therefore, shorten the assay readiness timeline.

Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta

“…The bridging ELISA is therefore often favored over the other assay formats. However, there are disadvantages with the bridging assay format since it detects disease-specific antibodies, such as RF that can bind to the drug causing interference (275). Direct ELISA, however, is even more prone to RF interference (87).…”

“…Direct ELISA, however, is even more prone to RF interference (87). Interference is an issue as it can confounds detection and interpretation of treatment-induced ADA (275). Moreover, the bridging assays cannot identify the IgG4 isotype (e.g.…”

Immunogenicity assay development and validation for biological therapy as exemplified by ustekinumab

“…It prevents p40 from binding to the receptor IL-12Rβ1 expressed on T cells. In summary, we present here different valid, easy-to-use and combinable approaches for assessing the immunogenic potential of a biotherapeutic, paving the way for a clinically relevant evaluation of immunogenicity and addressing challenges accompanied with it [4]. For the adequate testing of immunogenicity, the assessed methods were established or adapted to the requirements of the Food and Drug Administration (FDA) [2] and the European Medicines Agency (EMA) [3].…”

“…Neutralizing capacity of nADA was characterized using an adopted and validated cell-based assay. In summary, we present here different valid, easy-to-use and combinable approaches for assessing the immunogenic potential of a biotherapeutic, paving the way for a clinically relevant evaluation of immunogenicity and addressing challenges accompanied with it [4].…”

Development and characterization of an anti-rituximab monoclonal antibody panel

“…20 , 21 The strategy for immunogenicity assessment consists of several steps, including screening assay, confirmatory assay and characterization. In the screening and confirmation steps, ADAs with the potential to bind to the tested drugs are detected using various assay platforms, 22 , 23 including radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), electrochemiluminescence (ECL) immunoassay, surface plasmon resonance (SPR) assay and bio-layer interferometry (BLI) assay. Each of these binding assays has different sensitivity for the detection of ADAs in human clinical samples, including serum or plasma.…”