Enhancement of anti-thrombogenic properties of polypropylene (PP) to avert the adsorption of plasma proteins (fibrinogen and albumin), adhesion and activation of the platelets are very important for vast biomedical applications. The cold atmospheric pressure plasma (CAPP) assisted polymerization has potential to create the specific functional groups such as OCO, CO, CN and SS. on the surface of polymeric films using selective precursor in vapour phase to enhance anti-thrombogenic properties. Such functionalized polymeric surfaces would be suitable for various biomedical applications especially to improve the blood compatibility. The eventual aspiration of the present investigation is to develop the biofunctional coating onto the surface of PP films using acrylic acid (AAc) and polyethylene glycol (PEG) as a precursor in a vapour phase by incorporating specific functional groups for immobilization of biomolecules such as heparin (HEP), chitosan (CHI) and insulin (INS) on the surface of plasma modified PP films. The surface properties such as hydrophilicity, chemical composition, surface topography of the surface modified PP films were analyzed by contact angle (CA), Fourier transform infrared spectroscopy (FTIR), X-ray photo electron spectroscopy (XPS) and atomic force microscopy (AFM). Furthermore the anti-thrombogenic properties of the surface modified PP films were studied by in vitro tests which include platelet adhesion and protein adsorption analysis. It was found that the anti-thrombogenic properties of the PP films are effectively controlled by the CAPP grafting of AAc and PEG followed by immobilization of biomolecules of heparin, chitosan and insulin. The grafting and immobilization was confirmed by FTIR and XPS through the recognition of specific functional groups such as COOH, CO, SS and CN. on the surface of PP film. Furthermore, the surface morphology and hydrophilic nature of the PP films also tailored significantly by the successful grafting and immobilization which is confirmed by AFM and CA analysis. Owing to the physico-chemical changes on the surface of PP films induced by CAPP assisted polymerization, the anti-thrombogenic properties of PP films were enhanced as confirmed by in vitro analysis.