The stepwise construction of a novel kind of self-assembled organic/inorganic multilayers based on multivalent supramolecular interactions between guest-functionalized dendrimers and host-modified gold nanoparticles has been developed, yielding supramolecular layer-by-layer assembly. The deposition process was monitored by surface plasmon resonance spectroscopy. Further characterization of the multilayer films was performed by means of UV/vis absorption spectroscopy, which showed a linear increase in absorption with the number of bilayers. The growth of the gold nanoparticle plasmon absorption band corresponded to approximately a dense monolayer of gold nanoparticles per bilayer. Ellipsometry and atomic force microscopy (AFM) scratching experiments were used to measure the development of the film thickness with the number of bilayers, confirming linear growth and a thickness increase of approximately 2 nm/bilayer.