Neutron reflectivity measurements have been used to study the surface adsorption of the polyethylene sorbitan monostearate surfactant, with degrees of ethoxylation varying from 3 to 20 ethylene oxide groups, with the globular protein hydrophobin. The surface interaction between the ethoxylated polysorbate nonionic surfactants and the hydrophobin results in self-assembly at the air solution interface in the form of a well-defined layered surface structure. The surface interaction arises from a combination of the hydrophobic interaction between the surfactant alkyl chain and the hydrophobic patch on the surface of the hydrophobin, and the hydrophilic interaction between the ethoxylated sorbitan headgroup and the hydrophilic regions on the surface of the hydrophobin. The results presented show that varying the degree of ethoxylation of the polysorbate surfactant changes the interaction between the surfactant and the hydrophobin and the packing, and hence the evolution in the resulting surface structure. The optimal degree of ethoxylation for multilayer formation is over a broad range, from of order 6 to 17 ethylene oxide groups, and for degrees of ethoxylation of 3 and 20 only monolayer adsorption of either the surfactant or the hydrophobin is observed.