A bio-enabled method is presented to prepare a monolayers of titania nanoparticles using polyelectrolyte surface with tethered recombinant silaffin (rSilC). The surface-tethered silaffin molecules can be aggregated to produce TiO2 nanoparticles and to prevent the large-scale aggregation. The study used polyelectrolyte LbL films to control silaffin adsorption and enhance nanoparticles nucleation and growth. The study used LbL films made by polyelectrolytes, poly(allylamine hydrochloride) (PAH), and poly(styrene sulfonate) (PSS). The method successfully produced multilayer films with thickness from 4 to 20 nm. It was observed during the study that the confinement of titania nanoparticles into nanoscale protein surface domains can prevent the formation of larger microscopic aggregates.