Thromboembolism, arising from the utilization of cardiovascular medical devices, remains a prevalent issue entailing substantial morbidity and mortality. Despite the proposal of various surface modification strategies, each approach possesses inherent limitations and drawbacks. Herein, we propose a novel approach for the in situ growth of nanocoatings on various material surfaces through the cooperative assembly of silk fibroin (SF) and lysozyme. The intrinsic in situ growth characteristic enables the nanocoatings to achieve stable and uniform adherence to diverse substrate surfaces, including the inner surface of intravascular catheters, to redefine the surface properties of the material. The features of the hydrophilic and negatively charged nanocoating contribute to its antithrombotic properties, as evidenced by the reduced likelihood of platelet adhesion upon modification of the ultrathin and mechanically robust coating. In vitro assessment confirms a significant reduction in blood clot formation along with the promotion of anticoagulation. Such a SF/Ly nanocoating holds substantial promise as a surface modification strategy to enhance the hemocompatibility of medical devices and other materials that come into contact with blood, particularly in situations where medical-grade materials are temporarily unavailable, thus providing a feasible alternative.