Latex particle lithography is a highly reproducible method for patterning proteins using mild, ambient conditions and buffers. The natural self-assembly of monodisperse latex nanospheres on ultra-flat surfaces provides a structural template for patterning proteins such as bovine serum albumin (BSA). Solution mixtures of protein and polystyrene latex are drop-cast onto a flat substrate such as mica(0001) or Au(111). Monodisperse latex form colloidal crystalline arrays of close-packed structures on flat surfaces when dried. Proteins assemble in the uncovered surface areas between the latex spheres. After drying, the latex structural templates are removed by rinsing with water or buffers. The periodicity of protein nanostructures is determined by the latex diameter. However, we have also observed that the periodicity and morphology of the nanostructures is affected by the ratios of protein-to-latex. At certain ratios of protein it is more likely that ring structures will be formed. Typically, a layer of protein will be produced which is punctuated by pores or well-shaped structures where the latex was rinsed away. Ring structures are also observed, depending on experimental parameters. Thus, the diameter of the nanoparticles and the ratio of protein-to-latex determine the overall surface morphology of the patterned protein layer. Since this inexpensive and rapid method is capable of consistently producing large areas of protein nanostructures, it may be useful for biosensor surfaces in medical diagnostics and should also prove to be a valuable tool in investigating protein-antibody binding at the molecular scale.