The Research On The Preparation And Application Of The Fluorescent Protein Imprinted Materials

Proteomics is one of the most important research fields, which has a very closerelationship with human health and social development. The research on the protein isof great significance for the development of life science, which not only reveals liferule, but also provide the theoretical mechanism for understanding most diseases.However, various proteins always coexist in the real samples, for example, thecomponents in human and animal plasma is very complex. Therefore, selectiveseparation and identification of specific target protein from complex system hasbecome one of the bottle neck problems in protein research. Molecular imprintingtechnology (MIT) based on specific recognition ability for the target molecule hasbecome one of the ideal ways to solve this problem.In recent years, semiconductor quantum dots (QDs) have attracted great interestsdue to their extensive physical and chemical properties. If we combine the highselectivity of molecular imprinting technology and the excellent fluorescentcharacteristics of QDs, a new affinity material, with specific recognition cavity andresponding to the binding event with significant fluorescence intensity change, couldbe prepared. The most interest in the thesis is aimed at developing some novelmethods for the preparation of fluorescent protein imprinted polymers and theirapplication in selective separation of target protein.This thesis consists of five chapters. In Chapter1, the general introductionincluding the principle and historical retrospect of MIT and QDs, the application ofMIT and QDs in different fields, the development of new-style fluorescent molecularimprinting materials was described in details. Additionally, the aim and significanceof this thesis were also briefly presented.In Chaper2, a newly designed molecularly imprinted polymer (MIP) materialwas fabricated and successfully utilized as recognition element to develop QDs basedMIP-coated composite for selective recognition of the template cytochrome c. Thecomposites were synthesized by sol–gel reaction (imprinting process). The imprintingprocess resulted in an increased affinity of the composites toward the corresponding template. The fluorescence of MIP-coated QDs was stronger quenched by thetemplate versus that of non-imprinted polymer (NIP)-coated QDs, which indicatedthe composites could recognize the corresponding template. The results of specificexperiments further exhibited the recognition ability of the composites. The newcomposites integrated the high selectivity of MIT and fluorescence property of QDsand could convert the specific interactions between imprinted cavities andcorresponding template to the obvious changes of fluorescence signal.In Chaper3, a new type of MIP-based fluorescent artificial receptor wasdeveloped by anchoring MIP on the surface of denatured bovine serum albumin(dBSA) modified CdTe QDs using the surface molecular imprinting process. Theapproach combined the merits of molecular imprinting technology and the fluorescentproperty of the CdTe QDs. The dBSA was used not only to modify the surface defectsof the CdTe QDs, but also as assistant monomer to create effective recognition sites.Three different proteins, namely lysozyme, cytochrome c and methylated bovineserum albumin, were tested as the template molecules and then the receptors weresynthesized by sol–gel reaction (imprinting process). The results of fluorescence andbinding experiments demonstrated the recognition performance of the receptorstoward the corresponding template. Moreover, the new artificial receptors wereapplied to separate and detect lysozyme in real samples.In Chaper4, a facile approach was established for imprinting protein based onpolymerization of N-isopropylacrylamide (NIPAAm) and N,N′-methylenebisacrylamide (MBA) in the presence of template protein on thesilica-coated CdTe QDs surface. The silica coated on the surface of the QDs not onlyprotect luminescent QDs due to its optical transparence, but also provide the resultantcore-shell composite nanoparticles with good water dispersibility, biocompatibility,and surface functionality. The NIPAAm was introduced as temperature-sensitiveelement that allowed for swelling and shrinking in response to temperature changes torealize recognition and release of template BHb. Polymerization of NIPAAm andMBA can produce a thin MIP layer on the QDs surface. After removal of the templatemolecules, the imprinted sites were designed. The prepared MIP-based QDs exhibitspecific recognition behaviour to the corresponding template protein. In Chaper5, a new method for glycoprotein recognition was explored by usingaminophenylboronic acid functionalized mesoporous silica coating CdTe QDs andtheir combination with molecular imprinting. The preparation condition of fluorescentmaterial was investigated through the optimization of silane reagent, CTAB,phenylboronic acid, and so on. Then the structure and properties of mesoporousmaterials was charactered by the TEM, XPS and nitrogen adsorption. The selectiverecognition ability of the materials was exhibited through the UV-vis andfluorescence spectrophotometry, respectively, which showed that the material can beused for recognition of glycoproteins. Finally, molecularly imprinted material wasmodified on the surface of the aminophenylboronic acid functionalized mesoporoussilica coating CdTe QDs and the specific recognition ability of the resultant particleswas also explored.