Copolyamide films with a thickness from 50 to 780 nm were fabricated by interfacial polymerization between mixtures of m-phenylenediamine and primary amine-terminated polyamidoamine dendrimers (PAMAM) in the aqueous phase and trimesoyl chloride (TMC) in the organic phase. Different PAMAM generations (G0, d = 15 angstrom, Z = 4; G3, d = 36 angstrom, Z = 32; and G5, d = 54, Z = 128, where d is the measured diameter and Z is the number of terminal groups), and concentrations were used to obtain copolyamide films with different cross-linked structures. The influences of the concentration and degree of branching (PAMAM generation) on free volume were analyzed via positron annihilation spectroscopy (PAS) and correlated with the separation properties of copolyamide films. Besides, surface and intrinsic properties of copolyamide films prepared under different conditions were compared. The high hydrophilicity of PAMAM in the copolyamide network leads to the formation of a hydration layer on the copolyamide surface, which minimizes fouling. The separation performance of copolyamide membranes with various PAMAM networks was investigated in forward osmosis (FO) experiments. Understanding the correlation between the PAMAM structure/concentration, free volume, thickness, and surface intrinsic properties leads to the design of suitable fouling resistant thin-film composite membranes in a single interfacial polymerization process.