July 14-17, 2019
The formulation of therapeutic proteins is a critical process which aims at finding the most suitable conditions that impede protein degradation during long-term storage. One degradation path of high interest is the non-native aggregation1. The latter can be greatly suppressed by the selection of suitable solution conditions2. Over the years, various biophysical techniques have been explored as tools to quickly select the most promising formulations for long-term storage.
In this talk, we share the experience in our lab how some of these techniques can be integrated into protein formulation studies. We discuss the application of differential scanning calorimetry and the usefulness of determining the apparent protein melting temperatures to select protein formulations with high physical stability3. In the next part, we show how state-of-the-art equipment that employs fluorimetric and light scattering measurements during heating allows the sample saving characterization of proteins to select stable formulations. We then discuss the limitations of the existing non-isothermal techniques and show how the results from formulation studies can be biased when some excipient properties change during heating. In this context, we explore whether isothermal chemical denaturation is a useful complementary tool to overcome the limitations of non-isothermal techniques4.
Finally, we show how the assessment of the aggregation of partially folded species during refolding can provide additional information for the selection of protein formulations with high physical stability during storage4. The talk will be concluded with few general suggestions how to select solution conditions that impede aggregation during long-term storage of liquid protein formulations.
1. Roberts, C. J. Non-Native Protein Aggregation Kinetics. 98, (Springer New York, 2007).
2. Shire, S. J. Formulation and manufacturability of biologics. Curr. Opin. Biotechnol. 20, 708–714 (2009).
3. Youssef, A. M. K. & Winter, G. A critical evaluation of microcalorimetry as a predictive tool for long term stability of liquid protein formulations: Granulocyte Colony Stimulating Factor (GCSF). Eur. J. Pharm. Biopharm. 84, 145–155 (2013).
4. Svilenov, H., Markoja, U. & Winter, G. Isothermal chemical denaturation as a complementary tool to overcome limitations of thermal differential scanning fluorimetry in predicting physical stability of protein formulations. Eur. J. Pharm. Biopharm. 125, 106–113 (2018).
Hristo Svilenov and Gerhard Winter, "Biophysical characterization approaches to aid the selection of protein formulations by predicting their physical stability during long-term storage" in "Biological and Pharmaceutical Complex Fluids III: Protein Self-Assembly, Rheology and Interfacial Properties", Samiul Amin, Manhattan College, USA Miguel Rodrigues, University of Lisbon, Portugal Paolo Arosio, ETHZ, Switzerland Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/bpcf_iii/19