Layer-by-layer (LbL) assemblies have attracted much attention for their functional versatility and ease of fabrication. However, characterizing their thermal properties in relation to the film thickness has remained a challenging topic. We have investigated the role of film thickness on the glass transition temperature (T-g) and coeffecient of thermal expansion for poly(ethylene oxide)/poly(acrylic acid) (PEO/PAA) and PEO/poly(methacrylic acid) (PEO/PMAA) hydrogen-bonded LbL assemblies in both bulk and ultrathin films using modulated differential scanning calorimetry (modulated DSC) and temperature-controlled ellipsometry. In PEO/PAA LbL films, a single, well-defined T-g was observed regardless of film thickness. The T-g increased by 9 degrees C relative to the bulk T-g as film thickness decreased to 30 nm because of interactions between the film and its substrate. In contrast, PEO/PMAA LbL films show a single glass transition only after a thermal cross-linking step, which results in anhydride bonds between PMAA groups. The T-g, within error, was unaffected by film thickness, but PEO/PMAA LbL films of thicknesses below similar to 2.7 mu m exhibited a small amount of PEO crystallization and phase separation for the thermally cross-linked films. The coefficients of thermal expansion of both types of film increased with decreasing film thickness.