Novel methods for creating protein arrays with one and two-dimensional control can significantly enhance basic biological research as well as various bioarray applications. We demonstrate that structural variety and chemical heterogeneity of polystyrene-block-polymethylmethacrylate and polystyrene-b-poly(4-vinylpyridine) template surfaces can be successfully exploited to spontaneous formation of self-assembled, linear and hexagonally-ordered protein arrays that exhibit repeat spacings in a nanoscopic dimension. More importantly, protein molecules on the polymeric templates maintained their natural conformation and activity for several months. Our methods involving amphiphilic diblock copolymers to create highly-periodic, high density, surface-bound protein patterns also have the unique capability of controlling the exact repeat spacings of protein nanoarrays two-dimensionally by tuning the size of the underlying PS-b-PVP micellar templates. This additional advantage makes our platforms highly suitable as functional sensor substrates or protein arrays which demand precise spatial arrangements of protein molecules in multiple directions. Therefore, our novel protein assembly method will be greatly beneficial for high-throughput proteomic assays and multiplexed biosensing applications.