Synergistic recognition of transferrin by using performance dual epitope imprinted polymers

Transferrin (Trf) is a new type of active drug targeting carrier and disease biomarker that regulates the balance of iron ions in human body. The recognition and isolation of Trf is of great significance for disease diagnosis and treatment. Thus, a new type of magnetic dual affinity epitope molecularly imprinted polymer coated on Fe₃O₄ nanoparticles (Fe₃O₄@DEMIP) was successfully prepared for specific recognition of Trf. C-terminal nonapeptide and Trf glycan were selected as bi-epitope templates for metal chelation and boron affinity immobilization, respectively. 4-vinylphenylboric acid (4-VP), N-isopropyl acrylamide (NIPAM) and zinc acrylic were used as functional monomers. Results showed that Fe₃O₄@DEMIP exhibited excellent specific recognition ability adsorption capacity toward Trf, with an adsorption of 43.96 mg g^-1 (RSD = 3.28%) and a more satisfactory imprinting factor (about 6.60) than that of other reported imprinting methods. In addition, Fe₃O₄@DEMIP displayed pH, temperature and magnetic sensitivity properties to realize temperature and pH-controlled recognition and release of target proteins and magnetic rapid separation. Furthermore, the Fe₃O₄@DEMIP coupled with high-performance liquid chromatography (HPLC) analysis was successfully used for specific recognition of Trf in biosamples. This study provides a reliable protocol for preparing metal chelation and boron affinity dual affinity bi-epitope molecularly imprinted polymers for synergistic and efficient recognition of biomacromolecules in the complex biological systems.