文档介绍：河北大学
硕士学位论文
磁性微球的制备及其对BSA和酶的固定化研究
姓名:张亚芳
申请学位级别:硕士
专业:药物分析学
指导教师:张东浩
2011-06
摘要
摘要
磁性高分子微球是近年发展起来的一种新型功能高分子材料,它作为酶、细胞、药
物的载体被广泛地应用到细胞学、生物工程和生物医学等领域。特别是磁性微球用于固
定化酶技术,引起了研究者的极大关注。其中磁性高分子微球的结构及性能在很大程度
上直接影响着固定化蛋白或酶的催化活性及操作稳定性。所以载体的制备和表面性质将
成为固定化酶技术研究领域的重要内容。
本文在化学共沉淀法合成超细粉末Fe3O4的基础上,采用分散聚合法以苯乙烯(St)、
丙烯酰胺(AM)和丙烯酸(AA)为共聚单体,以过硫酸胺为引发剂(AIBN),乙醇/水混合溶
剂为分散介质,制备了微米级、单分散性较好的P(St/AM/AA)磁性高分子微球。对所制
备的磁流体和磁性微球的外观形态、粒径和表面羧基含量进行了表征。红外光谱表明St、
AM和AA在磁粒子表面发生了共聚合,扫描电镜的观察显示高分子微球的粒径为 µ
m- µm。
同时将不同单体比例制备的微球作为固定 BSA 的载体,考察了不同表面性能的磁
性微球对 BSA 固定化率的影响。结果表明,微球表面的亲疏水性不同,对 BSA 的固定
化率也不同。此外还研究了微球表面羧基含量,活化酯时间,固定化时间,温度和 pH
值对 BSA 固定化率的影响。在此基础上进一步研究磁性微球对 Candida rugosa 脂肪酶
(CRL)的固定化效果。结果表明,在固定了脂肪酶之后,磁性纳米微球仍具有优异的磁
分离能力。当固定化时间为 1 h 时,酶活达到 U/mg,酶活回收率达到 95 %。为深入
研究磁性微球固定化酶提供了有力的数据。
关键词磁粒子磁性微球牛血清白蛋白脂肪酶固定化率
I
Abstract
Abstract
ic polymer microsphere is a novel functional material that has been developed
resently. As the carrier of enzyme, cell and drug, it can be widely applied in many fields, such
as bio-engineering, cytology, biomedical science and so on. Especially, the application of
ic microspheres is of great interest for researchers in enzyme immobilization
technology. As an important part of immobilized enzymes, the structure and properties of
ic polymer microspheres affect the activities and operation stabilities of the
immobilized enzymes greatly. Therefore, the preparation of carrier materials as well as its
surface characteristics will be one of the most significant researches for the development of
immobilized enzymes in the future.
Here, the ic oxide (Fe3O4) was prepared by co-precipitation. Using ammonium
persulfate as initiator, ic functional polymer microsphere P(St/AM/AA) was
synthesized by the copolymerization of styrene, acrylamide and acrylic acid in aqueous
ethanol medium. In this paper, external shape, size, superficial acrylic acid content of the
microspheres and nano-particles of Fe3O4 were cha