血管抑素3A工程蛋白的柱复性与离子交换纯化

Separation of Anti-Angiogenic Agent from Inclusion Bodies by Column Refolding and Ion Exchange Adsorption

由大肠杆菌以包涵体形式表达的一种抗内皮细胞生长工程蛋白(A nti-ang iogen ic agent,简称3A)经变性,Sephacry l S-100 HR柱复性,SP Sepharose FF离子交换吸附纯化,SephadexG-25脱盐,获得复性率为53.47%,HPLC纯度为92.52%的3A活性蛋白。以猪髋动脉内皮细胞为受检细胞,表明纯化蛋白具有抑制内皮细胞生长的特性。

The isolation and purification of anti-angiogenic agent （3A）, a new functional iragment found in the tPA was studied. This process was started with the denaturation of anti-angiogenic agent protein expressed and accumulated in the the form of inclusion bodies in the cell of Escherichia coll. After column refolding using Sephacryl S-100 HR, SP Sepharose FF ion exchange adsorption, and Sephadex G-25 desalting, the active protein recovery and the purity of 3A determined by HPLC is 53.47% and 92.52% respectively. It was confirmed that the product protein can inhibit the growth of endothelioid cell.