Frozen state storage instability of a monoclonal antibody: aggregation as a consequence of trehalose crystallization and protein unfolding
| Autor: | Anjali P. Mehta, Steven C. Chico, Min Huang, Parag Kolhe, Satish K. Singh, Alanta L. Lary |
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| Rok vydání: | 2010 |
| Předmět: |
Protein Folding
Time Factors medicine.drug_class Drug Storage Pharmaceutical Science Monoclonal antibody Instability law.invention chemistry.chemical_compound Pharmaceutical technology Drug Stability law Freezing medicine Pharmacology (medical) Crystallization Protein Unfolding Pharmacology biology Chemistry Protein Stability Viscosity Organic Chemistry Antibodies Monoclonal Trehalose Biochemistry Immunoglobulin G Unfolded protein response biology.protein Molecular Medicine Thermodynamics Protein folding Antibody Biotechnology |
| Zdroj: | Pharmaceutical research. 28(4) |
| ISSN: | 1573-904X |
| Popis: | To investigate the cause of unexpected and erratic increase in aggregation during long-term storage of an IgG2 monoclonal antibody in a trehalose formulation at -20°C.Frozen matrix was sampled, stored frozen at various temperatures and analyzed by SEC over time.Aggregation increased with time at -20°C but not at -40°C or -10°C. The cause of the instability was the crystallization of freeze-concentrated trehalose from the frozen solute when the storage temperature exceeds the glass transition temperature of the matrix (-29°C). Crystallization at -20°C deprives the protein of the cryoprotectant, leading to a slow increase in aggregation. Storage at -10°C also leads to crystallization of trehalose but no increase in aggregation. It is hypothesized that significantly higher mobility in the matrix at -10°C allows protein molecules that are unfolded at the ice interface on freezing to refold back before significant aggregation can occur. In contrast, lack of mobility at -40°C prevents crystallization, refolding, and aggregation.Aggregation in the frozen state when stored above the glass transition temperature is a consequence of balance between rate of crystallization leading to loss of cryoprotectant, rate of aggregation of the unfolded protein molecules, and rate of refolding that prevents aggregation. |
| Databáze: | OpenAIRE |
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