A case study and use of sedimentation equilibrium analytical ultracentrifugation as a tool for biopharmaceutical development

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     

The effect of lactation length on the circulating concentrations of progesterone and oestradiol in the early weaned sow

Eighteen crossbred multiparous sows were allocated at random to one of two lactation lengths: 42 or 10 days. All sows were mated at the oestrus after weaning and from mating until day 26 post coitum they were bled every second day. Progesterone and oestradiol concentration in the plasma was determined by radioimmunoassay. Progesterone increased more rapidly between days 4 and 10 post coitum in early weaned sows and the oestradiol surge at mating was abnormally extended for the same group.