动态固定化方法制备微酶反应器研究初探

Preliminary Study on the Preparation of Enzyme Reactor using the Method of Dynamic Immobilization

以固定化人工膜为固定相,胰蛋白酶为固定靶蛋白,利用生物功能化色谱的方法,制备固定化微酶反应器,并与HPLC-MS/MS技术联用,检测目标蛋白牛血清白蛋白、肌红蛋白和细胞色素C的酶解效率。实验初步证明,所制备的新型微柱酶解器具有活性高、稳定性高、催化效率高、酶活性持久、快速再生及操作简单等优点。

In this study, an immobilized enzyme reactor was prepared in capillary by bio-functionalized chromatography method using immobilized artificial membrane as matrix and trypsin as immobilized enzyme. Bovine serum albumin, myoglobin, and cytochrome C were enzymolysised by the immobilized enzyme reactor and detected by HPLC-MS/ MS techniques. The preliminary results demonstrated that the activity, stability, and catalytic efficiency of the immobilized enzyme reactor were high. Moreover, its catalytic activity can be retained for a long time and the stationary phase can be reused.