Chemical in vitro models: assessing antibody quality attributes

Therapeutic antibody development traditionally relies on costly, time-consuming animal studies. This review examines how defined chemical models—including phosphate-buffered saline and continuous-flow dialysis systems—reliably predict antibody degradation, potentially reducing animal testing. By controlling physiological variables (pH, temperature, and redox potential), these models accurately predict pathways: deamidation, isomerization, pyroglutamate formation, thiol-disulfide exchange, and glycation. They are particularly valuable during early development with limited clinical material. Chemical in vitro approaches outperform traditional serum-based systems through superior control and reproducibility. We identify gaps, notably the lack of predictive oxidation models, and propose next-generation systems incorporating trace metals and antioxidants to simulate in vivo catalytic pathways. These advances align with the 3Rs principles (Replacement, Reduction, and Refinement) while accelerating development timelines and improving quality assurance.