Preparation And Application Of Molecular Imprinting Polymer In The Study Of Transferrin

Molecularly imprinted polymers(MIP)with specific binding sites show ultra-high selectivity to target molecules and are considered as synthetic antibodies.Surface imprinting technology is a method of generating imprinting sites on the surface of a substrate,which has been proposed to prepare protein or peptide imprinted polymers.Compared with traditional bulk imprinting,surface imprinting technology overcomes the difficulty of removing imprinted proteins from the polymer network.Glycoprotein-specific surface imprinted polymers(SMIP)have been widely reported.However,SMIP inevitably have disadvantages,such as insufficient recognition sites and low recognition performance.Therefore,the studies of increasing the specific surface area and the number of recognition sites of SMIP are very important to improve the capture efficiency of SMIP.The strategy of increasing specific binding sites through the multi-sites structure has received extensive attention.In order to further explore the application of MIP in the separation and recognition of macromolecular proteins,the main research contents of this article are listed as follows:(1)Using magnetic Fe₃O₄ nanoparticles as the carrier,glycans and polypeptides as the bi-epitope template,zinc acrylate,4-vinylphenylboronic acid,and N-isopropylacrylamide as functional monomers,a bi-epitope molecularly imprinted polymer Fe₃O₄@DEMIP that can specifically recognize the Trf was prepared.The experimental results showed that Fe₃O₄@DEMIP with high adsorption capacity and high imprinting factor has excellent specific recognition ability for Trf,the adsorption capacity and imprinting factor were 43.96 mg/g and 6.60,respectively.Adsorption experiments revealed that Fe₃O₄@DEMIP has p H,temperature and magnetic sensitivity characteristics,which can realize the identification and release of target proteins controlled by temperature and p H,as well as rapid magnetic separation.In addition,under optimal conditions,Fe₃O₄@DEMIP combined with high-performance liquid chromatography(HPLC)analysis has been successfully used for the specific identification of Trf in biological samples.It showed that Fe3O4@DEMIP can be used for the specific recognition and separation of biological macromolecules in complex biological systems.(2)By using the combined strategy of surface imprinting and epitope imprinting,with pyrrole and 3-amino benzene boric acid as functional monomer,in the presence of glycan and polypeptide double-epitope templates,a sensor GCE@MWCNTs-MIP,which can specific recognition of Trf,was prepared by electro-polymerization of molecularly imprinted polymer on multi-walled carbon nanotubes(MWCNTs)modified glassy carbon electrode(GCE)surface.Then,the sensor was used for the specific recognition and content determination of Trf in biological samples.The research results show that the combination of multi-walled carbon nanotubes modified glassy carbon electrode,bi-epitope template,pyrrole and 3-aminophenylboronic acid bifunctional monomer makes the prepared sensor have ultra-sensitive and selective recognition performance for Trf.The linear response range of the sensor to Trf is 0.1?M-1.25?M,the detection limit is 0.024?M(3?/S),which showed good stability,reproducibility and repeatability.In addition,the sensor has been successfully applied to the detection of Trf in serum samples.The spiked recovery rate is 94.70%?105.96%,and the RSD is2.64%?5.32%,indicating that the electrochemical sensor GCE@MWCNTs-MIP can be used for the detection of Trf in biological samples.