Protein stabilityEffect of hydration level on protein stability

Purpose and goal

The purpose of this project was to understand the relationship between the level of water content in solid formulations of therapeutic proteins and their thermal stability. To do so, we proposed two types of X-ray experiments (SAXS and W AXS) for the study of structural changes in a recombinant therapeutic protein during heat-induced denaturation at different moisture levels. By combining SAXS and WAXS, we planned to clarify the overall structural changes in thermal change. of proteins as well as their local structural Changes.

Expected results and effects

The studies have made it possible to investigate the relationship between protein structure and moisture in relation to thermal stability. The mechanism behind the stabilization of proteins with additives and the relation to the level of hydration is still debated after decades of research. It is well known that water content is important for the stability of dry protein drugs but not at what levels are critical and neither is the mechanism. Our new use of X-ray technology indicates opportunities for science-based development of long-lasting protein drugs.

Planned approach and implementation

The project activities consisted of producing series of protein formulations with certain moisture levels with subsequent X-ray studies of these formulations. The X-ray experiments have been performed at synchrotron plants in Hamburg and Barcelona in 2019. The results of these experiments have been evaluated in collaboration between Malmö Universitet, Sobi AB, RISE and MAX IV. The parties are already collaborating in other project which will strengthen the evaluation of new technology. Deeper analysis of data will need to be done and also require new processing methods