基于甲基丙烯酸接枝聚合的聚合物微球hemanvp对溶菌酶吸附及表面印迹的研究-study on adsorption and surface imprinting of lysozyme on polymer microsphere heman vp based on methacrylic acid graft polymerization.docx

在此适宜条件下，可制得接枝度为19.57g/100g的接枝微球PMAA-HEMA/NVP。以溶菌酶为碱性蛋白模型，深入研究了接枝微球PMAA-HEMA/NVP对碱性蛋 白的吸附性能与吸附机理。测定了接枝微球的zeta电位，考察了PMAA接枝度、介 质pH值及离子强度等因素对接枝微球PMAA-HEMA/NVP吸附性能的影响。研究结 果表明，在较大的pH范围内，接枝微球PMAA-HEMA/NVP的zeta电位为绝对值较 大的负值，即其表面携带有高密度的负电荷。在强静电相互作用的驱动下，接枝微 球PMAA-HEMA/NVP对溶菌酶表现出很强的吸附能力。随介质pH值的升高，接枝 微球对溶菌酶的吸附容量呈现先增大后减小的变化趋势，在与溶菌酶等电点接近的 pH值处（pH=9），具有最大的吸附容量（90mg/g）；离子强度对接枝微球的吸附能 力也有较大的影响，结果表明，当pH＜9时，溶菌酶吸附容量随 NaCl 浓度的增高 而减小；当pH＞9时，吸附容量随 NaCl 浓度的增高而增大。以改性微球 MAO-HEMA/NVP 为基质微球，以甲基丙烯酸为功能单体，N,N′- 亚甲基双丙烯酰胺为交联剂，以溶菌酶为模板分子，实施了溶菌酶分子的表面印迹， 制得了溶菌酶表面印迹材料 MIP-HEMA/NVP。以牛血红蛋白为对比物，考察了溶 菌酶印迹材料 MIP-HEMA/NVP 对溶菌酶的结合性能与识别特性。研究结果表明， 该印迹材料对溶菌酶具有特异的识别选择性与优良的结合亲和性，最高吸附量可达 到 216mg/g。相对于参比物牛血红蛋白，溶菌酶印迹材料对溶菌酶的吸附选择系数 为 31.070。此外，溶菌酶印迹材料也具有良好的解吸性能，以碳酸氢钠水溶液作为 洗脱液，洗脱效率可达 96.86%。在印迹过程中还发现，制备高结合选择性能溶菌 酶印迹材料的条件是：pH=9，交联剂用量与功能单体 MAA 量之比为 11.7:1。 关键词：生物相容性；聚合物接枝微球；甲基丙烯酸；碱性蛋白；溶菌酶；表面 印迹Studies on Adsorption and Surface Imprinting toward Lysozyme Based on Microsphere HEMA / NVP Grafting Polymerization of Methacrylic AcidAbstractHomopolymer of N-vinyl pyrrolidone (NVP) and hydroxyethyl methacrylate (HEMA) are polymer materials that both have good biocompatibility and even the blood compatibility, they both have importantly application in the field of biology and medicine. In this study, the functional microspheres PMAA-HEMA/NVP was obtained by grafting of polymethacrylic acid on cross-linked microsphere HEMA/NVP; then its properties and mechanism of adsorption for lysozyme that was the basic protein were explored; finally, the surface imprinting for lysozyme was studied on microsphere HEMA/NVP, and the imprinted materials MIP-HEMA/NVP that has a higher binding capacity and specific recognition properties for lysozyme was obtained. This study has important scientific significance and value in the field of Western blotting and biomedical materials.Firstly , the process conditions were improved about the preparation of cross-linked microsphere HEMA/NVP, The effects of various factors, such as the used amount of redistille