Abstract: Polydimethylsiloxane (PDMS) has good thermal stability, biocompatibility and corrosion resistance, and it has been widely used in the field of biomedical materials, i.e., medical catheters. However, the surface of PDMS is highly hydrophobic and susceptible to contamination by bacteria and plasma proteins. In this study, PDMS-TA-Ply surfaces were prepared by the rapid and successive deposition of tannic acid (TA) and ε-poly-L-lysine (Ply) on the PDMS surface via the solution immersion method. The surface chemical elements and hydrophilic properties of PDMS-TA-Ply surfaces were evaluated by X-ray photoelectron spectroscopy (XPS) and contact angle measurement. The anti-protein adsorption, antibacterial adhesion, anti-biofilm formation and cytotoxicity of PDMS-TA-Ply surfaces were evaluated. The results showed that PDMS-TA-Ply surfaces exhibited good antibacterial properties and low cytotoxicity toward L929 mouse fibroblasts. In addition, the PDMS-TA-Ply surface with higher content of Ply showed better antifouling/antibacterial performance.