Original scientific paper

https://doi.org/10.2478/acph-2024-0022

Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability

BLAŽ LEBAR ; University of Ljubljana, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, SI-1000 Ljubljana, Slovenia; Novartis, Global Drug Development, Technical Research & Development, Biologics, Drug Product Development, Formulation Development, Lek Pharmaceuticals d.d., SI-1000 Ljubljana, Slovenia
MITJA ZIDAR ; Novartis, Global Drug Development, Technical Research & Development, Biologics, Drug Product Development, Formulation Development, Lek Pharmaceuticals d.d., SI-1000 Ljubljana, Slovenia
JANEZ MRAVLJAK ; Novartis, Global Drug Development, Technical Research & Development, Biologics, Drug Product Development, Formulation Development, Lek Pharmaceuticals d.d., SI-1000 Ljubljana, Slovenia
ROMAN ŠINK ; Novartis, Global Drug Development, Technical Research & Development, Biologics, Drug Product Development, Formulation Development, Lek Pharmaceuticals d.d., SI-1000 Ljubljana, Slovenia
ALEŠ ŽULA ; University of Ljubljana, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, SI-1000 Ljubljana, Slovenia *
STANE PAJK ; University of Ljubljana, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, SI-1000 Ljubljana, Slovenia *

* Corresponding author.

Abstract

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 analyzing soluble aggregate formation through size exclusion chromatography. At low protein concentrations, protein instability after temperature stress was exclusively observed in the bis-TRIS/glucuronate buffer. Conversely, freeze-thaw stress led to a 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 concentrations, 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.

Keywords

biopharmaceuticals; buffer; stress; aggregates; alternative buffers; stability

Hrčak ID:

316645

URI

https://hrcak.srce.hr/316645

Publication date:

30.9.2024.

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