Abstract The antibacterial activity of magnetite-based nanocomposites synthesized by electron beam physical vapour deposition (EB PVD) was investigated. The phase, elemental and size composition of the synthesized nanoparticles were confirmed by scanning, transmission, X-ray and dynamic laser spectroscopy methods. The resulting EB PVD magnetite ligand-free nanoparticles (10.7 nm) were stabilized with polyvinylpyrrolidone and conjugated with the β-lactam antibiotic ampicillin, which increased the