The covalent attachment of proteins and their patterning on surfaces are important in the bio-medical industry, for use in applications such as bio-sensing and biocompatible medical devices. In this work, proteins are chemisorbed by reaction of their amine groups to glutaraldehyde- and carboxylic acid-functionalised surfaces, within patterns obtained by the dewetting of thin polystyrene films. The proof of principle is shown that consecutive attachment of proteins within the same pattern is possible using this method.