Nanostructured films consisting of polysaccharides and elastin-like recombinamers (ELRs) are fabricated in a layer-by-layer manner. A quartz-crystal microbalance with dissipation monitoring (QCM-D) is used to follow the build-up of hybrid films containing one polysaccharide (chitosan or alginate) and one of several ELRs that differ in terms of amino acid content, length and biofunctionality in situ at pH 4.0 and pH 5.5. The charge density of the ingredients at each pH is determined by measuring their ζ-potential, and the thickness of a total of 36 different films containing five bilayers is estimated using the Voigt-based viscoelastic model. A comparison of the values obtained reveals that thicker films can be obtained when working at a pH close to the acidity constant of the polysaccharide used (near-pKa conditions), suggesting that the construction of such films is more favorable when based on the presence of hydrophobic interactions between ELRs and partially neutralized polysaccharides. Further analysis shows that the molecular weight of the ELRs plays only a minor role in defining the growth tendency. When taken together, these results point to the most favorable conditions for constructing nanostructured films from natural and distinct recombinant polypeptides that can be tuned to exhibit specialized biofunctionality for tissue-engineering, drug-delivery and biotechnological applications.