【摘要】：蛋白質(zhì)是一種結(jié)構(gòu)復(fù)雜的生物大分子,具有良好的生物相容性,在生物醫(yī)學(xué)材料領(lǐng)域具有廣泛的應(yīng)用。但是,不同環(huán)境下的性質(zhì)差異如溶解性、保留時(shí)間、穩(wěn)定性等又限制了其進(jìn)一步的應(yīng)用,對(duì)蛋白質(zhì)進(jìn)行改性是拓寬其應(yīng)用范圍的有效方法之一。聚甲基丙烯酸甲酯(PMMA)是一種性能穩(wěn)定的疏水性材料。工業(yè)上通過單體的本體聚合和乳液聚合法制備PMMA,所得產(chǎn)物有良好的透光性和加工性能被稱為有機(jī)玻璃,并在涂料工業(yè)、光學(xué)材料等領(lǐng)域有廣泛應(yīng)用,同時(shí)PMMA有很好的生物相容性而被用于生物醫(yī)學(xué)等領(lǐng)域如作為血液透析的透析膜。因此將合成聚合物PMMA通過表面接枝(grafting to)的方式與蛋白質(zhì)結(jié)合制備兩親性蛋白質(zhì)-高分子雜化體,與傳統(tǒng)的兩親性嵌段共聚物相比,它不僅有自組裝特性更兼具生物相容性,有望在智能藥物控釋領(lǐng)域有良好的應(yīng)用。本文采用原子轉(zhuǎn)移自由基聚合(ATRP)法制備了分子量可控的端基官能化PMMA,考察了接枝牛血清白蛋白(BSA)合成兩親性BSA-PMMA雜化體的合成條件。通過核磁共振氫譜(1H-NMR)、凝膠滲透色譜(GPC)、變性蛋白質(zhì)聚丙烯酰胺凝膠電泳(SDS-PAGE)等方法對(duì)聚合物及雜化體結(jié)構(gòu)進(jìn)行了表征。隨后,采用動(dòng)態(tài)激光光散射(DLS)、掃描電子顯微鏡(SEM)、透射電子顯微鏡(TEM)等方法研究了BSA-PMMA雜化體在溶液中的的自組裝行為。首先,利用2-溴代異丁酸和N-羥基琥珀酰亞胺合成ATRP引發(fā)劑2-溴代異丁酸N-羥基琥珀酰亞胺酯(NHS-BIBA),以CuBr和N,N,N',N,'N''-五甲基二亞乙基三胺(PMDETA)為催化體系,以丙酮為溶劑,合成了端基官能化聚合物PMMA。通過1H-NMR和GPC對(duì)所得引發(fā)劑和聚合產(chǎn)物進(jìn)行表征,結(jié)果表明,成功合成了ATRP引發(fā)劑和分子量可控的端基官能化的PMMA。其次,研究了由親水的BSA與疏水的PMMA制備具有兩親性的BSA-PMMA雜化體,確定了構(gòu)建兩親性BSA-PMMA雜化體的最佳條件。通過變性蛋白聚丙烯酰氨凝膠電泳(SDS-PAGE)測(cè)試的結(jié)果表明,在丙酮:水=1:25,pH=5的條件下PMMA能夠很好地通過表面接枝的方式與BSA結(jié)合形成雜化體。最后,通過激光動(dòng)態(tài)散射儀(DLS)、掃描電子顯微鏡(SEM)、透射電子顯微鏡(TEM)研究了兩親性BSA-PMMA雜化體在溶液中的自組裝行為。DLS測(cè)試的結(jié)果表明,兩親性BSA-PMMA雜化體在溶液中的自組裝形成了膠束,隨著雜化體中聚合物分子量或比例的增加自組裝膠束粒徑增大,膠束溶液的Zeta電勢(shì)結(jié)果為-44.5mv,表明溶液穩(wěn)定性很好。SEM測(cè)試的結(jié)果表明,兩親性BSA-PMMA雜化體自組裝形成各種復(fù)雜的空間結(jié)構(gòu)如囊泡結(jié)構(gòu)(直徑約500nm)、棒狀結(jié)構(gòu)、膜狀結(jié)構(gòu)等。TEM測(cè)試的結(jié)果表明,兩親性BSA-PMMA雜化體自組裝形成的膠束外壁具有流動(dòng)性。
[Abstract]:Protein is a complex biomolecules with good biocompatibility and is widely used in biomedical materials. However, the differences of properties in different environments, such as solubility, retention time, stability and so on, restrict its further application. Modification of protein is one of the effective methods to broaden its application range. Polymethyl methacrylate (PMMA) is a stable hydrophobic material. In industry, the products obtained by bulk polymerization and emulsion polymerization of monomers have good transmittance and processability, and have been widely used in coating industry, optical materials and other fields. At the same time, PMMA has good biocompatibility and has been used in biomedical fields such as hemodialysis as dialysis membrane. Therefore, amphiphilic protein-polymer hybrids were prepared by surface grafting of PMMA with protein. Compared with traditional amphiphilic block copolymers, it not only has self-assembly properties, but also has biocompatibility. It is expected to have a good application in the field of intelligent drug controlled release. Amphiphilic BSA-PMMA hybrids were prepared by atom transfer radical polymerization (ATRP) and their molecular weight controlled terminal functionalized PMMA, (PMMA,) were prepared. The synthesis conditions of amphiphilic BSA-PMMA hybrids were investigated by grafted bovine serum albumin (BSA) (BSA). The structures of polymers and hybrids were characterized by 1H-NMR and (GPC), denatured protein gel electrophoresis (SDS-PAGE). Then the self-assembly behavior of BSA-PMMA hybrids in solution was studied by means of dynamic laser light scattering (DLS), scanning electron microscope (SEM), transmission electron microscope (TEM) and so on. Firstly, 2-bromoisobutyric acid and N-hydroxysuccinimide were used to synthesize 2-bromoisobutyric acid N-hydroxysuccinimide (NHS-BIBA) initiator. The terminal functionalized polymer (PMMA.) was synthesized by using 'Nishi-pentamethylenediethylenetriamine (PMDETA) as catalyst system and acetone as solvent. The initiators and polymerization products were characterized by 1H-NMR and GPC. The results showed that ATRP initiator and PMMA. with controllable molecular weight were successfully synthesized. Secondly, the preparation of amphiphilic BSA-PMMA hybrids from hydrophilic BSA and hydrophobic PMMA was studied, and the optimal conditions for constructing amphiphilic BSA-PMMA hybrids were determined. The results of polyacrylamide gel electrophoresis (SDS-PAGE) of denatured protein showed that under the condition of acetone: water = 1: 25 (pH = 5), PMMA could bind to BSA by surface grafting to form hybrids. Finally, the self-assembly behavior of amphiphilic BSA-PMMA hybrids in solution was studied by (DLS), scanning electron microscope (SEM),) and transmission electron microscope (TEM) with a dynamic laser scattering instrument. The self-assembly of amphiphilic BSA-PMMA hybrids in solution formed micelles. With the increase of molecular weight or proportion of polymer in the hybrid, the particle size of self-assembled micelles increased, and the Zeta potential of micellar solution was -44.5 MV. The results of SEM test show that the amphiphilic BSA-PMMA hybrids self-assemble into various complex spatial structures, such as vesicle structure (diameter about 500nm), rod structure, membrane structure, etc. The results of TEM test show that, The outer wall of micelle formed by self-assembly of amphiphilic BSA-PMMA hybrids has fluidity.
【學(xué)位授予單位】：蘭州交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：O631.1;O629.73

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本文編號(hào)：2328340