Argon plasma-pretreated polytetrafluoroethylene (PTFE) films were subjected to further surface modification by near-UV light-induced graft copolymerization with acrylic acid (AAc), sodium salt of styrenesulfonic acid (NaSS), and N,N-dimethylacrylamide (DMAA). The surface compositions and microstructures of the modified films were characterized by angle-resolved X-ray photoelectron spectroscopy (XPS). A stratified surface microstructure with a significantly higher substrate-to-graft chain ratio in the top surface layer than in the subsurface layer was always obtained for PTFE surface with a substantial amount of the hydrophilic graft. The stratified surface microstructure was consistent with the observed hysteresis in advancing and receding water contact angles. The graft yield increased with Ar plasma pretreatment time and monomer concentration. Covalent immobilization of trypsin on the AAc polymer-grafted PTFE films was facilitated by water-soluble carbodiimide(WSC). The effective enzyme activities increased initially with increasing surface concentration of the grafted AAc polymer but became saturated at a moderate AAc polymer concentration. The immobilized enzyme could still retain close to 30% of its original activity. Solution-coating of the polymeric acid-modified PTFE films with the emeraldine (EM) base of polyaniline readily resulted in an interfacial charge transfer interaction and a semiconductive PTFE surface.