In this paper, we report the controlled fabrication of layer-by-layer (LbL) films deposited on gold substrates with three different supramolecular architectures using polypyrrole (Ppy) and magnetite nanoparticles (Fe 3 O 4-np), besides conventional poly(allylamine hydrochloride) (PAH) e poly(vinyl sulfonic acid) (PVS) polyelectrolytes, demonstrating the synergistic effect between Ppy and Fe 3 O 4-np such as a result of their interaction. Modified gold electrodes were analyzed by contact angle (wettability), surface plasmon resonance (SPR), raman spectroscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. The (Fe 3 O 4-np/Ppy) 3 architecture was also evaluated by scanning electron microscopy. The modified gold electrodes present more homogeneous covering, higher electron transfer and a decrease of resistance with the incorporation of the nanostructured materials such as Ppy and Fe 3 O 4-np forming (Fe 3 O 4-np/Ppy) 3 LbL film. The results carried out in this study suggest that the (Fe 3 O 4-np/Ppy) 3 LbL film can be applied as a possible electrochemical or optical non-enzymatic sensor for analytical detection.