Physicochemical analysis and biological characterization of FKB327 as a biosimilar to adalimumab

FKB327 was approved by the European Medicines Agency as a biosimilar to European-authorized adalimumab (Humira® ; AbbVie Inc). Adalimumab is a monoclonal antibody, binding and inhibiting tumor necrosis factor (TNF)-α with use indicated for several immune-mediated, chronic, and inflammatory disorders. The approval is based on high similarity in the physicochemical properties between FKB327 and adalimumab. The objective of this study is to assess the biological similarity, with regard to Fab- and Fc-associated functions, and describe the relationship between physicochemical and biological characterization and functional activity. State-of-the-art orthogonal techniques were implemented to assess the structure and function of FKB327. Peptide mapping with liquid chromatography and mass spectrometry, capillary electrophoresis-sodium dodecyl sulfate, ultraviolet circular dichroism, size-exclusion high-performance liquid chromatography (HPLC), and cation exchange HPLC were the techniques used to assess structure. Functional activity was assessed with enzyme-linked immunosorbent assay, surface plasmon resonance, and cell-based assays. The polypeptide sequence of FKB327 was identical to that of adalimumab. FKB327 also was demonstrated to have a similar secondary and tertiary structure to adalimumab. Posttranslational heterogeneities, along with size and charge variants, were not clinically meaningful. FKB327 binds to TNF-α, FcγR, the neonatal Fc receptor, and C1q, and induces apoptosis, antibody-dependent cellular cytotoxicity, and complement-dependent cytotoxicity. The binding and activity of FKB327 were similar to that of adalimumab. FKB327 shares similar structure and activity with adalimumab. Based on characterization of physicochemical and biological properties, FKB327 is expected to have a similar safety, immunogenicity, and efficacy profile to adalimumab