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 l...
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2024-09-01
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| Series: | Acta Pharmaceutica |
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| Online Access: | https://doi.org/10.2478/acph-2024-0022 |
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| author | Lebar Blaž Zidar Mitja Mravljak Janez Šink Roman Žula Aleš Pajk Stane |
| author_facet | Lebar Blaž Zidar Mitja Mravljak Janez Šink Roman Žula Aleš Pajk Stane |
| author_sort | Lebar Blaž |
| collection | DOAJ |
| description | 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 histi-dine/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. |
| format | Article |
| id | doaj-art-f62cd29e0f684705b3c342fb13655542 |
| institution | Kabale University |
| issn | 1846-9558 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Sciendo |
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| series | Acta Pharmaceutica |
| spelling | doaj-art-f62cd29e0f684705b3c342fb136555422025-02-02T08:34:38ZengSciendoActa Pharmaceutica1846-95582024-09-0174347949310.2478/acph-2024-0022Alternative buffer systems in biopharmaceutical formulations and their effect on protein stabilityLebar Blaž0Zidar Mitja1Mravljak Janez2Šink Roman3Žula Aleš4Pajk Stane5University of Ljubljana Faculty of Pharmacy, Department of Pharmaceutical Chemistry, SI-1000Ljubljana SloveniaNovartis, Global Drug Development, Technical Research & Development Biologics, Drug Product Development, Formulation Development Lek Pharmaceuticals d.d.SI-1000Ljubljana, SloveniaUniversity of Ljubljana Faculty of Pharmacy, Department of Pharmaceutical Chemistry, SI-1000Ljubljana SloveniaNovartis, Global Drug Development, Technical Research & Development Biologics, Drug Product Development, Formulation Development Lek Pharmaceuticals d.d.SI-1000Ljubljana, SloveniaNovartis, Global Drug Development, Technical Research & Development Biologics, Drug Product Development, Formulation Development Lek Pharmaceuticals d.d.SI-1000Ljubljana, SloveniaUniversity of Ljubljana Faculty of Pharmacy, Department of Pharmaceutical Chemistry, SI-1000Ljubljana SloveniaThe 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 histi-dine/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.https://doi.org/10.2478/acph-2024-0022biopharmaceuticalsbufferstressaggregatesalternative buffersstability |
| spellingShingle | Lebar Blaž Zidar Mitja Mravljak Janez Šink Roman Žula Aleš Pajk Stane Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability Acta Pharmaceutica biopharmaceuticals buffer stress aggregates alternative buffers stability |
| title | Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability |
| title_full | Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability |
| title_fullStr | Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability |
| title_full_unstemmed | Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability |
| title_short | Alternative buffer systems in biopharmaceutical formulations and their effect on protein stability |
| title_sort | alternative buffer systems in biopharmaceutical formulations and their effect on protein stability |
| topic | biopharmaceuticals buffer stress aggregates alternative buffers stability |
| url | https://doi.org/10.2478/acph-2024-0022 |
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