Adaptation of an ELISA assay for detection of IgG 2a responses against therapeutic monoclonal antibodies in a mouse immunization model.
Shraddha S RaneRebecca J DearmanIan KimberJeremy P DerrickPublished in: Journal of immunotoxicology (2022)
Biotherapeutic monoclonal antibodies (mAb) play important roles in clinical medicine but their potential to elicit immune responses in patients remains a major issue. In a study designed to investigate the effect of aggregation on immunogenic responses, mice were immunized with two monoclonal antibodies (mAb1 and mAb2). Serum levels of total IgG, IgG 1 , and IgG 2a were measured by ELISA. An anti-mouse IgG 2a monoclonal detection antibody cross-reacted with mAb2 but not mAb1, leading to high background when the ELISA plate was coated with mAb2. The problem was solved by use of a goat anti-mouse IgG 2a polyclonal antibody that demonstrated the required specificity. IgG 2a responses were similar for monomer- or aggregate-coated ELISA plates. The results demonstrate the importance of assessment of the specificity of individual reagents when measuring antibody responses against therapeutic antibodies by ELISA.