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Study On The Synthesis And Application Of Novel Biomaterials

Posted on:2004-01-18Degree:DoctorType:Dissertation
Country:ChinaCandidate:H H YangFull Text:PDF
GTID:1101360092470996Subject:Uncategorised
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This dissertation consists of five chapters, including the overview of intelligent biomaterials, sol-gel biomaterials and nanometer fluorescence probes; synthesis and application of novel environment sensitive polymers; synthesis and application of novel sol-gel biomaterial; synthesis and application of nanometer fluorescence hybrid silica particles; the conclusion and prospect of the present research work.In chapter one, highlights of new findings in biomaterials' studies and its significance in modern biological engineering and medicine engineering are introduced. The mechanisms, synthesis methods, characters studies and biological applications of environment sensitive polymers are viewed. The brief introduction and current development of sol-gel biomaterials are given. The use of nanometer particles as biological labeling is summarized. The research proposal for this dissertation is also presented.In chapter two, the research work on the synthesis and biological applications of the novel environment sensitive polymers is described. The results enlarge the application range of environment sensitive polymers and have positive significance for developing the application of environment sensitive polymers in analytical biochemistry and biological engineering. This chapter is divided into two sections.In section one, an oligo-N-isopropylacrylamide (ONIPAAm) with a terminal carboxyl group is prepared by polymerization of NIPAAm with mercaptopropionic acid. The ONIPAAm exhibits a low critical solution temperature of about 32-33 癈. This oligomer is then conjugated to antibody, oligonucleotides and mimeticAbstractenzyme by using N-(3-Dimethylaminopropyl)-Af'-ethylcarbodhydro-chlorid (EDCI) as a coupling reagent. (1) Because the antibody is covalently immobilized to the very end of ONIPAAm chains, the antibody aggregation is eliminated and the steric hindrance that occurs in the free radical copolymerization of antibody and poly-N-isopropylacrylamide is also decreased. Therefore, better bioactivity of the immune complex is retained. The immune activity of immobile antibody and non-specific adsorption of antibody and antigen on the ONIPAAm chains are studied. These characters are employed to develop a novel thermally separated fluorescence immunoassay method for the determination of mouse IgG. Under optimum conditions, the calibration graph for the mouse IgG is linear over the range of 0.5-1000 ng/mL, with a detection limit of 0.2 ng/mL (3a/S). (2) The study discovers that the conjugate between ONIPAAm and (dT)8 maintains a high affinity with its complementary sequence (dA)8 and can be applied to the specific separation of (dA)g form its mixed solution with (dA)3dT(dA)4. Then the ONIPAAm-(dT)8-(dA)g solution can be heated to the melting temperature of the duplex between (dT)8 and (dA)s to get the purified (dA)8. This method can be used for the separation and detection of polyadenylated mRNAs which are of great importance in cDNA cloning. (3) Our experiment shows that the conjugate between ONIPAAm and iron-tetrasulfonatophthalocyanine (FeTSPc) displays high peroxidatic activity for the fluorogenic reaction of p-HPPA and H2O2. The Kcat ratio of ONIPAAm-FeTSPc to free FeTSPc is about 6:1, namely, the catalytic activity of FeTSPc increases by more than six times when FeTSPc is bound to the oligomer. The possibility of its application in analytical field is also tested by the determination of H2O2 and ONIPAAm-FeTSPc, the detection limits for H2O2 and ONIPAAm-FeTSPc are 4.2 x 10'9 and 1.7x10-9mol L-1, respectively.In section two, the synthetic method and analytical application of temperature-and pH-sensitive polymers are given. A series of pH-sensitive phase separating polymers containing both temperature- and pH-sensitive components are synthesized by copolymerizing N-isopropylacrylamide and methacrylic acid (MAA). A P(NIPAAM-co-MAA) polymer is finally obtained, with a lower critical solution pH (LCSP) about 5.8 at 37C. The r-values of NIPAAm and MAA are 0.862...
Keywords/Search Tags:biomaterials science, environment sensitive polymers, sol-gel, magnetic particles, nanometer fluorescence probe, enzyme catalysis, fluorescence immunoassay, flow injection immunoassay
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