A case study and use of sedimentation equilibrium analytical ultracentrifugation as a tool for biopharmaceutical development
| Autor: | A. J. Brown, N. R. Burns, H. C. Dawkes, P. G. Varley |
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| Rok vydání: | 1997 |
| Předmět: |
Chromatography
Chemistry Osmolar Concentration Size-exclusion chromatography Biophysics A protein Saccharomyces cerevisiae General Medicine Macrophage Inflammatory Proteins Recombinant Proteins Biopharmaceutics Molecular Weight Analytical Ultracentrifugation Biopharmaceutical Drug Design Sedimentation equilibrium Chromatography Gel Mutagenesis Site-Directed Cloning Molecular Chemokine CCL4 Ultracentrifugation Chemokine CCL3 Protein Binding |
| Zdroj: | European Biophysics Journal. 25:437-443 |
| ISSN: | 1432-1017 0175-7571 |
| DOI: | 10.1007/s002490050058 |
| Popis: | Analytical ultracentrifugation (AUC) has reemerged as a powerful technique for protein characterisation. We report the pivotal role sedimentation equilibrium AUC has played in the development of macrophage inflammatory protein-1 alpha (MIP-1 alpha) as a protein therapeutic. MIP-1 alpha has potential clinical applications in cancer but its clinical use is limited, since it associates to form large insoluble aggregates in physiological buffers. Using AUC as a screening technique, we have produced a biologically active variant of MIP-1 alpha, BB-10010, which has a reduced tendency to aggregate in physiological buffers. The aggregation of protein based pharmaceuticals is routinely monitored by size exclusion chromatography (SEC). Comparison of the data acquired by SEC and AUC, demonstrates that owing to the complexity of BB-10010, AUC analysis is required in addition to SEC to provide a rigorous characterisation of molecular association. This work has been extended to include the use of AUC as an analytical tool to monitor the quality of BB-10010 during formulation and stability studies. |
| Databáze: | OpenAIRE |
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