In the present study, three thermoplastic polyurethanes (TPUs) based on poly(tetramethylene glycol) (PTMG) and poly(butylene adipate) (PBA) as polyols as well as their nanocomposites containing graphene oxide GO (0.3–1%) were prepared. TPUs had T_gs and T_ms values in the range of −62.5 to −21.6°C and 5 to 10.8°C, respectively, increasing with enhancing PBA content in their formulations. Increasing of PBA moieties in TPUs resulted in higher crystallinity, elastic modulus and tensile strength values, while elongation at break and Helium permeability were decreased. Incorporation of GO into the TPUs improved crystallinity, mechanical properties and gas barrier values.