Synthesis and functionalization of scalable and versatile 2D protein films via amyloid-like aggregation

Two-dimensional (2D) protein films can be used to modify the properties of surfaces, and find applications predominantly in the fields of biomaterials, lithography, optics and electronics. However, it is difficult to produce scalable homogeneous and robust protein films with an easy, low-cost, green and efficient method. Further challenges include encapsulating and releasing functional building blocks in the film without inactivating them, and maintaining or improving the bioactivities of proteins used for the formation of the films. Here we detail the process to prepare large 2D protein films with user-defined features and structures via the amyloid-like aggregation of commonly synthesized proteins. These films can be synthesized at meter scales, have high interface adhesion, high functional expansibility and tunable functional properties, obtained by controlling the position of the disulfide bond breakage. For example, we can retain or even enhance the natural antibacterial, biomineralization and antifouling activity of proteins involved in film formation, and the properties can also be expanded through the physical blending or chemical grafting of additional functional blocks on the surface of the film. A 2D protein film can be prepared in ~3 h using four alternative coating techniques: immersion, transfer, hydrogel stamping and spraying. The characterization process of the film requires ~5 d. The procedure can be carried out by users with basic expertise in materials science.