琼脂糖微球的制备及活化条件研究[精选].doc

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、NaBH4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%，NaBH4的浓度为0.5g/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 Abstract Generally speaking, the higher the activation degree of agarose magnetic microspheres is, the more affinity ligands can be 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 magnetic 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，NaBH4 concentration，activation temperature and activation 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 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 magnetic 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 目录 摘要 I Abstract II 目录 III 第一章 文献综述 1 1