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Implant-associated infection is one of the most common and problematic complications for dental and orthopaedic patients. Modification of currently used implant surfaces aimed at bestowing them with antibacterial properties is a promising approach in the development of new biomaterials. In this study, a novel nanotechnology, that is, electrostatic self-assembly, was developed to construct biomimetic polypeptide nanocoatings on commonly used metal implants. A model antibacterial drug, cefazolin, was captured in the polypeptide nanocoating and its release was studied. We have shown that the capture and release of cefazolin was pH-induced and could be controlled, and the developed antibiotic-incorporated polypeptide multilayer nanocoatings could prevent Staphylococcus aureus colonization thus showing great potential for preventing implant-associated infection.