Development of multifunctional, bio‐active surfaces has become a major focus of biomedical engineering in the last decade. By using physicochemical surface modification technique, it is possible to equip biomedical surfaces with numerous properties, including enhanced cell proliferation and adhesion, as well as antibacterial or anti‐inflammatory effects. Principally, this has been achieved through surface functionalization with biochemistries to elicit specific biological functions. With respect to biochemical functionalization, both the substrate and the immobilization method can affect the performance of immobilized biomolecules, influencing their stability and activity. Covalent immobilization, particularly, is a favorable strategy to form long‐lasting biomedical coatings with versatile biological activity stable over time. This review combines an overview of current and emerging immobilization techniques coupled with an in‐depth investigation of the underlying mechanisms governing the activity and stability of covalently immobilized biomolecules. By dissecting the intricate interplay between immobilized biomolecules, cellular interactions, and functional outcomes, this study addresses a pivotal knowledge gap and presents an indispensable guide for the development of tailored biofunctionalized surfaces in the realm of biomedical engineering.