This work presents the development and optimization of two fabrication methods that combine plasma polymerization and electron beam lithography techniques for producing chemical patterns at the sub-micro scale. The first method uses sacrificial resist as mask to produce the functionalization of the bioadhesive areas and the second method consists in the direct EBL writing of adhesive regions on a non-adhesive plasma deposited PEO-like film. The produced patterned surfaces exhibit high binding capacity as tested by Surface Plasmon Resonance Imaging with Human Serum Albumin. Furthermore, we show that adsorption on the structured surfaces is similar to that observed on a flat surface with a direct proportionality to the active area. This work shows the efficiency and suitability of the developed chemical patterning techniques and their high potential applicability for the study of protein surface interactions and miniaturized biosensing device development.