Structural stability of the template is one of the most important considerations during the preparation of protein imprinting technology. To address this limitation, we propose a novel and versatile strategy of utilizing macromolecularly functional monomers to imprint biomacromolecules. Results from circular dichroism and synchronous fluorescence experiments reflect the macromolecularly functional monomers tendency to interact with the protein surface instead of permeating it and destroying the hydrogen bonds that maintain the protein's structural stability, therefore stabilizing the template protein structure during the preparation of imprinted polymers. The imprinted polymers composed of macromolecularly functional monomers or their equivalent micromolecularly functional monomers over silica nanoparticles were characterized and carried out in batch rebinding test and competitive adsorption experiments. In batch rebinding test, the imprinted particles prepared with macromolecularly functional monomers exhibited an imprinting factor of 5.8 compared to those prepared by micromolecularly functional monomers with the imprinting factor of 3.4. The selective and competitive adsorption experiments also demonstrated the imprinted particles made by macromolecularly functional monomers possessed much better selectivity and specific recognition ability for template protein. Therefore, using macromolecularly functional monomers to imprint may overcome the mutability of biomacromolecule typically observed during the preparation of imprinted polymers, and thus promote the further development of imprinting technology.