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

The isolation and purification of anti-angiogenic agent(3A),a new functional fragment 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 coli.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.