A biosensor calibration structure is provided that includes at least two electrode structures in which at least one of the electrode structures has a non-random nanopattern on the sensing surface which provides a different sensing surface area than at least one other electrode structure. The at least one other electrode structure may be non-patterned (i.e., flat) or have another non-random nanopattern on the sensing surface. A biological functionalization material such as, for example, glucose oxidase or glucose dehydrogenase, can be located on at least the sensing surface of each electrode structure. The biosensor calibration structure can be used within a biosensor calibration method.