Development of Relative Potency Assay for ADC Using Multiple Kinetic Assays

Developing relative potency cell-based assays (CBAs) for antibody-drug conjugates (ADCs) involves several key steps and considerations. CBA for ADC usually assesses ADC induced cytotoxicity through endpoint viability readout, for example, quantification of ATP level as indicator of metabolically active cells with CellTiter-Glo® Cell Viability Assay (Promega). Monitoring ADC-induced apoptosis and secondary necrosis before assessing endpoint viability could provide a more comprehensive understanding of the ADC's effects on cells addressing some of the challenges associated with endpoint viability assays, such as confirmation of an ADC’s mechanism of action (MoA), cell plating density optimization, ADC dose-response curve and appropriate endpoint timing selection. By incorporating apoptosis and secondary necrosis monitoring, you might gain insights into the timing and progression of cell death, which could help refine your endpoint viability assessments.