| Molecularly imprinted technique?MIT?is developed to obtain the molecularly imprinted polymers?MIPs?that match the templates in their shape,size and functional groups.In recent years,with the rapid development of MIT,MIPs have been successfully used in the recognition and separation of amino acids,saccharides,sugar derivatives and drugs.However,due to the complex configurations of the biomolecules such as peptides,proteins,enzymes,nucleic acids and DNA,there is a vital problem of MIPs for the recognition and separation for a long time.Moreover,with the further research of MIT,some scholars have found that it was hard to elute and transfer the templates for the MIPs that were prepared using embedding method.All these problems have severely limited the application and development of MIT.Therefore,it is extremely promising to explore a novel approach for the MIPs to recognize and separate the biomolecules effectively.Our research was aimed to resolve these problems in consideration of all these existing problems.In order to effectively solve the recognition and separation for immunostimulating peptide of human,a novel method has been proposed using ionic liquid functionalized nano/microspheres as carrier and theoretical calculation,mainly in the following areas.?1?Surface molecularly imprinted nano/microsphere?MIMs?was successfully prepared by the surface grafting polymerization method with thymopentin?TP5?as template through ionic liquid-functionalized polyethyleneglycolmethacrylateco-vinylimidazole microsphere as the matrix.The surface structure and property of MIMs have been fully characterized by SEM,FTIR and XPS.The results showed that the maximum adsorption capacity of MIMS for TP5was 38.4 mg·g-1 with the IF of 1.96 under the concentration of TP5 with 0.2 mg·mL-1,indicating that MIMs showed good specific property for TP5.The specific property of MIMs was due to the fact that there were matched imprinting sites between the imidazolium and carboxyl groups of MIMs and TP5 through the non-covalent interactions.?2?Novel water-compatible ionic liquid-functionalized microspheres?HCCSMIMs?with molecularly imprinted shell layer were controllably synthesized via precipitation polymerization and surface imprinting technique.Here,a room-temperature ionic liquid was synthesized to prepare these surface-imprinted microspheres with excellent water solubility and multiple binding sites with template molecules.Results reveal that ionic liquid incorporation significantly improves the adsorption of TP5.Moreover,the adsorption property and recognition capability towards TP5 are closely related to the synergetic effect of electrostatic interaction and hydrogen bonding.Through employing the synergetic effect of directional and non-directional interactions,the surface-imprinted microspheres exhibit high adsorption capacity,good selective recognition,and rapid binding ability for TP5.The surface-imprinted microspheres demonstrate potential usage for TP5 enrichment from other biomolecules,and the proposed method was successfully applied for TP5 determination in thymopentin injection and urine.?3?The surface functionalities of nanomaterials provide an invaluable resource to biomaterials science in the form of biomolecular recognition,sensing,self-assembly and drug delivery.This paper presents a onestep procedure for the facile preparation of highly cross-linked raspberry-like nano/microspheres by the dispersion polymerization of polyethylene glycol dimethacrylate and N-vinylimidazole and studied the mechanism of preparation in detail.Then,ionic liquid-functionalized reactions were conducted on the surface of the raspberry-like nano/microspheres to investigate the effects of the functional groups on the peptide adsorption in aqueous media.The results reveal that the nano/microspheres were raspberry-like,monodisperse and stable.It was also found that the adsorption of immunostimulating hexapeptide from human?IHH?was correlated with the surface properties of the ionic liquid-functionalized nano/microspheres,which demonstrated their potential use for the immobilization and release of IHH in practical application.In addition,theoretical calculation was used to investigate the binding mechanism of raspberry–like nano/microspheres with highly cross–linked structure for binding at the molecular level.Moreover,the interactions between nano/microspheres and IHH have been investigated at the molecular level.The overall properties of nano/microspheres could be reflected by studying their chemical and geometrical properties of functional chains.This work not only could provide theoretical basis for designing nano/microspheres with specific surface properties with widespread application,but also provide theoretical and experimental basis for MIPs with specific recognition properties with precise controlling.Particularlly,this work has laid the foundation for the following research with further mechanism investigation of molecular imprinting.?4?In order to further investigate the mechanism of molecularly imprinting,surface-imprinted microspheres MRHCMIMs were controllably synthesized using ionic liquid-functionalized microspheres with a highly cross-linked raspberry-like structure as the matrix via surface molecular self-assembly and precipitation polymerization in aqueous media at room temperature.Results reveal that the adsorption capacity and selective recognition of MRHCMIMs have a direct relationship with the properties of the functional chain of the ionic liquid-functionalized microspheres.The mechanisms of the template immobilization and molecularly imprinting have been fully presented.Moreover,MRHCMIMs that have both high adsorption capacity and good selective recognition were used in competitive rebinding tests and analysis of urine samples,which demonstrated their potential use for IHH enrichment and in real samples. |
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