The interface molecules designed to exhibit molecular recognitions between different species have become attractive tools for the bottom-up fabrication and hybridization of nanostructured units. Here, we focus on antibodies with high binding ability and specificity to construct a novel biomolecule interface for recognizing an inorganic material. Careful selection from a phage-displayed library of variable region heavy and light Fv chains of human antibodies using enzyme-linked immunosorbent assay and surface plasmon resonance assay resulted in the identification of an antibody fragment, A14P-b2, with high affinity (KD = 1.7 nm) and specificity for gold materials. Our results indicated the potential usefulness of human antibody libraries and the effectiveness of the antibody framework for recognizing bulk material surfaces. Construction of bivalent and bispecific antibodies on the A14P-b2 platform with high affinity by means of fusion technology enabled the functionalization of gold nanoparticles and allowed selective protein accumulation on gold spots patterned on a silicon substrate. This type of antibody engineering is potentially applicable to bio-inspired materials and nanobiosensing.