Aims: The applicability of particle lithography with monodisperse mesospheres is tested with various proteins to control the surface coverage and dimensions of protein nanopatterns. Methods & Materials: The natural self-assembly of monodisperse spheres provides an efficient, high-throughput route to prepare protein nanopatterns. Mesospheres assemble spontaneously into organized crystalline layers when dried on flat substrates, which supply a structural frame or template to direct the placement of proteins. The template particles are displaced with a simple rinsing step to disclose periodic arrays of protein nanopatterns on surfaces. Results & Discussion: The proteins are attached securely to the surface, forming nanopatterns with a measured thickness of a single layer. The morphology and diameter of the protein nanostructures can be tailored by selecting the diameter of the mesospheres and choosing the protein concentration. Conclusions: Particle lithography is shown to be a practical, highly reproducible method for patterning proteins on surfaces of mice, glass and gold. High-throughput patterning was achieved with ferritin, apoferritin, bovine serum albumin and immunoglobulin-G. Depending on the ratio of proteins to mesospheres, either porous films or ring structures were produced. This approach can be applied for fundamental investigations of protein-binding interactions of biological systems in surface-bound bioassays and biosensor surfaces.
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