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1、广西大学本科毕业论文 琼脂糖磁性微球活化条件研究- I -摘要一般说来，琼脂糖磁性微球活化度越高，能连接的亲和配位体就越多，用其吸附纯化蛋白质及固定化酶效率就越高。本实验研究了用环氧氯丙烷活化琼脂糖磁性微球的最优条件。实验先用反相悬浮包埋法制备出琼脂糖磁性微球，微球因内部含有磁性四氧化三铁的超细粉末而具有磁响应性。以微球表面的环氧基密度作为活化度的度量，考察了 NaOH 浓度、环氧氯丙烷体积分数、NaBH 4 的浓度、活化温度和活化时间等因素对琼脂糖磁性微球活化反应活化度的影响，得到了最佳活化条件为：NaOH 溶液浓度为0.9mol/L，环氧氯丙烷体积分数为 40%，NaBH 4 的浓度为 0
2、.5g/L，活化温度为 40，活化时间为 4h。用环氧氯丙烷活化的载体稳定性好，且可提供相当于 3 个碳原子间距的空间臂。本文还研究过相似活化步骤和条件下，改用 1,4-丁二醇二缩水甘油醚做活化剂，但未能成功活化琼脂糖磁性微球，该研究尚待进一步开展。关键词：琼脂糖 磁性微球 活化 广西大学本科毕业论文 琼脂糖磁性微球活化条件研究- II -AbstractGenerally speaking, the higher the activation degree of agarose magnetic microspheres is, the more affinity ligands can b
3、e grafted. So that the adsorptive capacity of microspheres for protein purification and the efficiency of immobilized enzyme can be higher. This paper investigated optimal conditions for the activation of agarose magnetic microspheres, using epichlorohydrin as activating agent. Firstly, agarose magn
4、etic microspheres were prepared by reverse suspended embedding method. The microspheres possessed magnetic responsibility because of the interior Fe3O4 ultrafine powder. The effects of sodium hydroxide concentration，epichlorohydrin volume fraction，NaBH 4 concentration，activation temperature and acti
5、vation time on activation degree were investigated，according to the density of surface epoxy group . The optimal conditions are obtained as followed：sodium hydroxide concentration，epichlorohydrin volume fraction， NaBH4 concentration ，activation temperature and activation time is 0.9 mol/L, 40%, 0.5
6、g/L, 40 , 4 h, respectively. The carrier activated by epichlorohydrin can be of good stability and has 3 carbon atom space arm. This paper also studied the activation of agarose magnetic microspheres using 1, 4-butyl glycol two glycidyl ether as activating agent. The experiments in which agarose mag
7、netic microspheres were failed to be activated were operated under similar methods and conditions. The relevant researches are yet to be further developed.Keywords：Agarose; magnetic microspheres; activation广西大学本科毕业论文 琼脂糖磁性微球活化条件研究- III -目录摘要 .IABSTRACT .II目录 .III第一章 文献综述 .11.1 固定化酶技术的研究意义 .11.2 固定化酶
8、载体基质材料 .21.2.1 固定化酶载体基质材料性质要求 .21.2.2 固定化酶载体材料研究进展 .21.3 固定化酶载体的表面修饰 .31.3.1 固定化酶载体表面修饰原则 .31.3.2 固定化酶载体表面修饰环氧基 .41.4 固定化酶载体的间隔臂 .61.5 固定化酶载体的配位体 .71.6 展望 .71.7 本论文研究内容及意义 .7第二章 琼脂糖微球的制备 .92.1 实验试剂及仪器 .92.1.1 实验试剂 .92.1.2 实验仪器 .92.2 实验方法及步骤 .102.3 实验结果及讨论 .10第三章 琼脂糖磁性微球活化条件研究 .123.1 微球活化方法论证 .123.2 环氧基分析方法论证 .123.3 实验试剂及仪器 .13广西大学本科毕业论文 琼脂糖磁性微球活化条件研究- IV -3.3.1 实验试剂 .133.3.2 实验仪器 .143.4 实验方法及步骤 .
