Harnessing Multi-Anchoring Effects for the Fabrication and Specific Recognition of Surface-Oriented Imprinted Nanospheres for Cytochrome C

Protein molecularly imprinted polymers (MIPs), as artificial antibodies, are promising for protein separation due to their low cost, easy preparation, and high stability, but their performance is limited by poor mass transfer, imprecise imprinting, and single interaction modes. Herein, dendritic mesoporous silica nanoparticles (DMSNs) were used as the support, and a self-designed multifunctional poly(ionic liquid) macromonomer (p(VIMCD-co-VAIM-co-VSIM-co-VVIM)) served as the functional monomer to achieve directional anchoring of cytochrome C (Cyt-C). Surface-imprinted microspheres (DMSNs@MPS@PILs-MIPs) were prepared via free-radical copolymerization for Cyt-C recognition. The DMSNs possessed interconnected mesoporous channels, good dispersibility, an average particle size of ~80 nm, and a specific surface area of 267.97 m2/g. Ionic liquid monomers were synthesized via alkylation, and the macromonomer was constructed through a two-step method. Molecular dynamics simulations and spectroscopic characterization revealed the macromonomer-stabilized Cyt-C conformation, with interactions dominated by van der Waals forces. The DMSNs@MPS@PILs-MIPs featured a thin imprinted layer (~5 nm) to reduce mass-transfer resistance. Adsorption studies showed Cyt-C adsorption followed Langmuir and pseudo-second-order models, with a maximum capacity of 383.14 mg/g and an imprinting factor of 2.17. Only 12% capacity loss occurred after repeated cycles, indicating robust regeneration stability. This study provides a feasible strategy for constructing protein surface-imprinted polymers based on multifunctional synergistic interactions and conformational stabilization.