In recent years, gold nanoparticles have been utilized in biomedical applications due to their biocompatibility as well as unique optical properties. In order for the gold nanoparticles to become effective agents, they are functionalized with vectors that will target the nanoparticles to specific cells to be imaged. Conventionally, hybrid gold nanostructures have been produced by complex, multistep chemical cross-linking bioconjugation procedures usually involving biologically-unfriendly stringent conditions. To overcome difficulties related to conventional cross-linking methods, we propose a novel surface functionalization platform using biocompatible gold-binding peptide templates (AuBP). AuBPs were previously selected from combinatorial FliTrx peptide library and demonstrated nanomolar binding affinities to a gold surface under physiological conditions. We then conjugated AuBP with other binding domains engineered to bind to the constant Fc-region of an antibody (FcBP). This antibody is then recognized by receptors in the cell membrane for absorption. The bifunctional AuBP-FcBP peptides self-assemble on the surface of the gold nanoparticles, and effective gold binding is shown through a localized surface plasmon resonance, high-salinity stability assay and gel electrophoresis. A dot blotting assay is used to confirm the binding of the antibodies. These peptide-functionalized and cell-specific nanoprobes have a great potential for developing novel bioimaging tools for diagnostics and therapy.