Hydrophobin protiens are unique to the fungal kingdome and have evolved to function in different roles during the growth and development of filamentous fungi. Due to the unique properties of these proteins to self-assemble into amphipathic monolayers at hydrophobic:hydrophlic interfaces, they can be found as biosurfactants, protective coatings and as primers to enhance surface adhesion. In recent decades there has been a significant development towards the applying these proteins to a range of different research fields, from food technology and surface coatings to drug delivery devices. However, the understanding of the mechanism in which these proteins undergo self-assemble at the interface is still lacking. In this project, I have combined high resolution imagaing techniques, such as AFM, TEM and TEM tomography to compare the substructure of different Class I hydrophobin rodlet films, and using colometric kinetic assays to delineate a model for the assembly mechanisms at the interface. With the information, I was able to reveal that the exposure of hydrophobin proteins to the surface interface is a determining factor. By altering the surface interface with additives, such as ethanol, it was possible to manipulate the hydrophobin film structure and physioproperties. This knowledge was used to successfully formulate a nanosupsension of hydrophobin with hydrophobic compounds, such as curcumin and Amphotericin B. This research project lays the foundation for the future development and refinement of hydrophobin-based technologies.