The Influence of Arginine and Counter-Ions: Antibody Stability during Freeze-Drying

Abstract

Amino acids, for example L-arginine, are used in lyophilisation as crystalline bulking, buffering, viscosity reducing or stabilising excipients. In this study, arginine was formulated with different counter ions (hydrochloride, citrate, lactobionate, phosphate, and succinate). A monoclonal antibody was investigated in sugar-free arginine formulations and mixtures with sucrose regarding cake appearance and protein aggregation and fragmentation. Arginine hydrochloride formulations collapsed during lyophilisation due to its low Tg' and partially crystallised during storage, but provided the best protein stability at low antibody concentration, followed by arginine succinate. Arginine citrate/phosphate/lactobionate formulations resulted in amorphous elegant cakes, but inferior protein stability. Addition of sucrose improved cake appearance and protein stability. Arginine phosphate with sucrose resulted in similar protein stability as the sucrose reference. Mixtures of sucrose with arginine hydrochloride/lactobionate/succinate provided better stability than sucrose alone. While 50 mg/mL antibody improved the cake appearance, only arginine lactobionate provided sufficient protein stability next to sucrose. Overall, sugar-free arginine hydrochloride and lactobionate lyophilisates stabilised the antibody comparably or better than sucrose depending on antibody concentration. The best protein stability was found for mixtures of arginine hydrochloride/lactobionate/succinate with sucrose.