Preparation And Application Of Protein Molecularly Imprinted Materials

Protein molecularly imprinted materials have attracted much attention due to their great potential in biotechnology and biomedical engineering.Although molecular imprinting techniques are widely used in small molecules for chemistry and biomedicine,imprinting of biomacromolecules,especially proteins,remains a significant challenge.In recent years,many protein molecularly imprinted materials have been successfully prepared and used in chromatographic separation,biosensing and drug delivery design.These materials are low cost and easy for preparation,have good biocompatibility and special ability for recognition of proteins.However,it also has some shortcomings,such as low template utilization and non-biocompatible preparation conditions.This dissertation mainly focuses on the application of protein molecularly imprinted materials in adsorbent materials,fluorescence detection and gene delivery,and the main works are as follows:(1)In this work,we prepared a magnetic molecularly imprinted nanoparticles(DES-MIPs)with a deep eutectic solvent(DES)as a functional monomer to separate transferrin(TrF)in human serum.The DES and TrF are combined by electrostatic interaction,and the imprinted cavities can specifically adsorb TrF through electrostatic action and configuration for improving selectivity.The DES dosage,pH and time for adsorption have been optimized,and this materials showed special adsorption property for TrF.The prepared material provides a biocompatible and time-saving method for the isolation and purification of proteins.(2)A magnetic fluorescent molecularly imprinted nanoparticles(FMINPs)was prepared for extraction and fluorescence detection of TrF.The FMINPs was prepared with two steps,the first step was the synthesis of magnetic TrF imprinted nanoparticle and the second step was introducing a near-infrared fluorescent compound(CyA)on the imprinted nanoparticles,which has a strong near infrared fluorescence emission at730 nm while excitation at 690 nm and a large fluorescence signal quenching after adsorption of TrF.The concentration of TrF can be determined by the change of the fluorescence signal.This method can be directly used to separate and detect proteinsin samples,and successfully applied to the detection of TrF concentration in human serum samples.(3)Based on the low biocompatibility of protein molecularly imprinted materials,the DES was used as a biocompatible functional monomer to prepare a fluorescent molecularly imprinted nanogel for separation and detection of myoglobin.After electrostatic adsorption CyA,the entire material has a strong fluorescence emission.Cytochrome c(Cyt C)in myoglobin can cause intense quenching of CyA.By using this mechanism,the myoglobin is detected.The fluorescent molecularly imprinted nanogel has the potential to be applied to detect myoglobin in vivo due to its good biocompatibility and small particle size,and successfully applied to detect myoglobin in human serum samples.(4)For widespread the application of protein molecularly imprinted materials in treatment of cancers and early diagnosis.We developed a kind of high-loading molecularly imprinted gelatin nanoparticles(GDMINPs)to deliver DNA probe to cancer region,detect and image telomerase in situ.When the protein molecularly imprinted material enters the body,it will adsorb TrF in situ to form a protein crown,which can successfully resist the immune stress and allow the gene delivery obtained the longer circulation time.The results show that the material can be successfully applied to in situ imaging of telomerase in mice tumor models,and provide an empirical set of data for designing future generations of gelatin materials for DNA delivery.