Antifouling coatings stand out as one of the highly efficient ways to mitigate surface contamination. Traditional antifouling coatings have a major drawback: they rely on highly toxic and environmentally hazardous compounds. These substances not only lead to ecological harm but also disrupt the natural equilibrium of ecosystems. Consequently, in recent years, eco-friendly antifouling bio-coatings have emerged. This review focuses on the mechanisms and processes underlying contaminant adhesion, laying a solid foundation for grasping the principles of antifouling coating design. It further elaborates on the general strategies for developing bio-based antifouling solutions, highlighting their potential across a wide array of applications. Finally, this review carefully analyzes the current challenges confronted by antifouling bio-coatings and puts forward future development directions. Through a comprehensive overview, we aim to expand the influence of bio-based antifouling technologies, promote the further application of bio-based antifouling coatings in marine antifouling and medical antifouling fields, and provide examples for the establishment of environmental protection policies.