20.500.12556/RUL-163471 Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability The formulation of biopharmaceutical drugs is designed to eliminate chemical instabilities, increase conformational and colloidal stability of proteins, and optimize interfacial stability. Among the various excipients involved, buffer composition plays a pivotal role. However, conventional buffers like histidine and phosphate buffers may not always be the optimal choice for all monoclonal antibodies (mAbs). In this study, we investigated the effects of several alternative buffer systems on seven different mAbs, exploring various combinations of ionic strengths, concentrations of the main buffer component, mAb concentrations and stress conditions. Protein stability was assessed by analysing soluble aggregate formation through size exclusion chromatography. At low protein concentration, protein instability after temperature stress was exclusively observed in the bis-TRIS/glucuronate buffer. Conversely, freeze-thaw stress led to significant increase in aggregate formation in tested formulations, highlighting the efficacy of several alternative buffers, particularly arginine/citrate, in preserving protein stability. Under temperature stress, the introduction of arginine to histidine buffer systems provided additional stabilization, while the addition of lysine resulted in protein destabilization. Similarly, the incorporation of arginine into histidine/HCl buffer further enhanced protein stability during freeze thaw cycles. At high protein concentration, the histidine/citrate buffer emerged as one of the most optimal choices for addressing temperature and light induced stress. The efficacy of histidine buffers in combating light stress might be attributed to the light absorbing properties of histidine molecules. Our findings demonstrate that the development of biopharmaceutical formulations should not be confined to conventional buffer systems, as numerous alternative options exhibit comparable or even superior performance. biopharmaceuticals buffer stress aggregates alternative buffers stability biofarmacevtika pufer stres agregati alternativni pufri stabilnost farmacevtska kemija true false true Angleški jezik Slovenski jezik Članek v reviji 2024-10-07 12:41:38 2024-10-07 12:41:42 2024-10-10 03:35:17 0000-00-00 00:00:00 2024 0 0 Str. 479–493 no. 3 Vol. 74 2024 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 615.4:54 1846-9558 10.2478/acph-2024-0022 197188099 Alternative-buffer-systems-in-biopharmaceutical-formulations-and-their-effect-on-protein-stability.pdf Alternative-buffer-systems-in-biopharmaceutical-formulations-and-their-effect-on-protein-stability.pdf 1 D188B32023A1AE46771A0EA4C73199F0 5ffae56d3b56cafe0ae159accc23468a96902d2efeb6ed0260a5dfe6de0ee131 3e598527-8499-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=193183 https://acta.pharmaceutica.farmaceut.org/wp-content/uploads/2024/09/47924.pdf 1 9252155f-8498-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=193182 https://sciendo.com/article/10.2478/acph-2024-0022?tab=abstract 1 92331f8c-8498-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=193181 Fakulteta za farmacijo 0 0 0