大肠杆菌表达的重组白细胞介素-2的初步纯化

Preliminary Purification of Recombinant Interleukin-2 Expressed in E. coli

本文对大肠杆菌表达产生的重组白细胞介素-2进行了纯化研究。通过比较两种方法制备的rIL-2包含体的纯度,发现用4mol/L脲溶解可溶性细菌蛋白后可使rIL-2包含体纯度达70％;在高浓度变性剂条件下进行凝胶过滤,解决了rIL-2易聚合的问题;结合透析,利用空气氧化形成高活性氧化型rIL-2;经SephadexG-100凝胶过滤,DEAE离子交换等步骤纯化,得到了均一性rIL-2,纯度达98％,比活达4.3×10~6u/mg蛋白,得率为30.8％。

This Paper studied the purification of rIL-2 expressed in E.coli.By Comparing the purity of rIL-2 inclusion bodies produced in two different ways,we found that the purity of rIL-2 IBs may reach to 70% by dissolving the soludble bacterium proteins with 4mol/L urea.The polymerization of rIL-2 had been prevented by gel filtration in the presence of high concentration denaturant.The highly active oxy rIL-2 was formed by oxidizationin air during dialysis.After Sephadex G-100 gel filtration and DEAE ion exchange,homogeneous rIL-2 of 98% purity and 30.8% of recovery was obtained.The Specific actively was 4.3**********x106U/mg.