Optimization of Streptokinase Mutant Protein Purification Method Using Affinity Chromatography Technique

Abstract

Protein purification has always been one of the most critical and challenging stages of drug-protein production. Streptokinase as the most common, and currently, the most cost-effective fibrinolytic drug is no exception. In this study, the mutated streptokinase producing clone (SK263cyc) to which the histidine tag was added was grown in TY2x medium, and SDS-PAGE assessed protein expression after induction of protein expression. Three different methods did protein purification; in the first one, metal, ion affinity chromatography (IMAC) technique was used. In the second solution, first, by filtration with ammonium sulfate, the purification was carried out, and then by affinity purification, chromatography continued. In the third solution, hydrophobic chromatography was used to purify the streptokinase protein. The purity of ophthalmic purity was 93.2%, and the purity of hydrophobic purity was about 90.4%, whereas the combination of pre-treatment with ammonium sulfate and the purity of the ophthalmic method did not achieve more than 88%.In general, the results of this study show that the IMAC method is more suitable as a final method in the process of streptokinase purification than the other two approaches.