A case study and use of sedimentation equilibrium analytical ultracentrifugation as a tool for biopharmaceutical development |
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Authors: | P G Varley A J Brown H C Dawkes N R Burns |
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Institution: | (1) British Biotech Pharmaceuticals Ltd., Watlington Road, Oxford, OX4 5LY, UK (Fax: +44-1865-781187), GB |
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Abstract: | Analytical ultracentrifugation (AUC) has re-emerged as a powerful technique for protein characterisation. We report the pivotal
role sedimentation equilibrium AUC has played in the development of macrophage inflammatory protein-1α (MIP-1α) as a protein therapeutic. MIP-1α 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-1α, 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.
Accepted: 6 October 1996 |
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Keywords: | Analytical ultracentrifugation Biopharmaceutical Protein Chemokine |
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