The functionalization of polytetrafluoroethylene (PTFE) for human thrombomodulin (hTM) binding has been achieved by CO2 plasma activation and subsequent vapour phase graft polymerization of acrylic acid (AA). The PTFE surfaces after CO2 plasma treatment, AA grafting and hTM immobilization were characterized by attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS) and electron spin resonance (ESR) spectroscopy, as well as by zeta potential and wetting measurements to quantitatively control each step of modification. The activity of immobilized hTM was estimated by the protein C activation test.