A new approach is introduced to create submicrometer patterned surfaces using multilayer polymer films that contain alternating layers of two polymers, linear low-density polyethylene (LLDPE) and ethylene-co-(acrylic acid) copolymer (EAA). Patterned templates have been prepared by microtoming the multilayer molded sheets. Regionally confined chemical functionality is confirmed by grafting an amine-terminated biotin and adsorbing streptavidin specifically on the alternating layers of EAA.