The main aim of this investigation was to study the interaction of sulfated xylans as antithrombotic substances with poly(ethylene terephthalate) (PET) model films as a model for blood contacting surfaces. The adsorption of sulfated xylans onto PET model films was studied as a function of pH and ionic strength using the quartz crystal microbalance with dissipation (QCM-D) technique. The application of positively charged polyethyleneimine (PEI) as an anchoring polymer was done to improve the adsorption. The hydrophilic/hydrophobic properties of functionalized PET surfaces were monitored by goniometry, whilst their elemental composition was determined by X-ray photoelectron spectroscopy. Sulfated xylans adsorbed favorably at pH 5 by physical interactions and by entropy gain driven adsorption. Higher ionic strengths of solutions improved adsorption, due to the reduction of electrostatic repulsive forces between PET surfaces and anionic xylans' macromolecules. The intermediate PEI layer caused more extensive and stable adsorption due to Coulomb interactions. The surface modifications presented in this work provided important information regarding the adsorption/desorption phenomena between antithrombotic sulfated xylans and PET surfaces. The latter is of great interest when preparing advanced polymer composite material such as functional antithrombotic PET surfaces for blood-contacting medical devices and presents an extremely challenging research field.