The effect of mAb and excipient cryoconcentration on long-term frozen storage stability - part 2: Aggregate formation and oxidation
| Autor: | Oliver Bluemel, Jakob W. Buecheler, Astrid Hauptmann, Georg Hoelzl, Karoline Bechtold-Peters, Wolfgang Friess |
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| Rok vydání: | 2021 |
| Předmět: |
frozen storage
SEC Size-exclusion chromatography cryoconcentration PES Polyethersulfone Pharmaceutical Science HPLC High performance liquid chromatography stability FCM Freeze-concentrated matrix HPW Highly purified water Tg′ Glass transition temperature of the maximally freeze-concentrated solution RS1-441 OD350 Optical density at 350 nm Pharmacy and materia medica monoclonal antibody DLS Dynamic light scattering SVP Subvisible particle mAb Monoclonal antibody large-scale freezing HMWS Higher molecular weight species Research Paper |
| Zdroj: | International Journal of Pharmaceutics: X International Journal of Pharmaceutics: X, Vol 4, Iss, Pp 100109-(2022) |
| ISSN: | 2590-1567 |
| Popis: | We examined the impact of monoclonal antibody (mAb) and buffer concentration, mimicking the cryoconcentration found upon freezing in a 2 L bottle, on mAb stability during frozen storage. Upon cryoconcentration, larger protein molecules and small excipient molecules freeze-concentrate differently, resulting in different protein to stabiliser ratios within a container. Understanding the impact of these shifted ratios on protein stability is essential. For two mAbs a set of samples with constant mAb (5 mg/mL) or buffer concentration (medium histidine/adipic acid) was prepared and stored for 6 months at −10 °C. Stability was evaluated via size-exclusion chromatography, flow imaging microscopy, UV/Vis spectroscopy at 350 nm, and protein A chromatography. Dynamic light scattering was used to determine kD values. Soluble aggregate levels were unaffected by mAb concentration, but increased with histidine concentration. No trend in optical density could be identified. In contrast, increasing mAb or buffer concentration facilitated the formation of subvisible particles. A trend towards attractive protein-protein interactions was seen with higher ionic strength. MAb oxidation levels were negatively affected by increasing histidine concentration, but became less with higher mAb concentration. Small changes in mAb and buffer composition had a significant impact on stability during six-month frozen storage. Thus, preventing cryoconcentration effects in larger freezing containers may improve long-term stability. Graphical abstract Unlabelled Image |
| Databáze: | OpenAIRE |
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