Room Temperature Intrinsic Emission Ratio of BSA Correlates With Percent Aggregates During Long-Term Storage.

Successful formulation development hinges on the ability to screen and identify excipients that stabilize drug products during long-term storage. Biophysical and accelerated stability studies are used to screen for excipients that stabilize protein drug products. However, these studies are not always predictive of aggregation during long-term storage. In this study, we used multivariate experimentation to compare the effectiveness of intrinsic fluorescence and size exclusion chromatography accelerated stability parameters to predict excipients that stabilized bovine serum albumin (BSA) against aggregation on long-term storage at 4 °C. Emission intensity ratio (IR) data was more sensitive than emission maxima (λ) or intensity measurements in identifying significant factors and interactions. We observed the expected inverse correlation between the mid-points of fluorescence thermal transitions (Ts) and insoluble aggregates at 4 and 40 °C. However, there were positive correlations between Ts and % aggregates at 4 °C, indicating that if T was used as a predictive tool, it would select formulations that promoted soluble aggregates on long-term storage. Ambient temperature IR measurements identified excipients that reduced BSA soluble aggregates on long-term storage. The results show ambient temperature emission ratio measurements can be useful for protein formulation development.