Optimization of Streptokinase Mutant Protein Purification Method Using Affinity Chromatography Technique

Authors

  • Nastaran Rahimi1 , Mohammad AlinezhadChamazketi2, Amir Yaghoubi Nezhad3, Forough Talaeizadeh4

DOI:

https://doi.org/10.37506/ijfmt.v14i2.3383

Keywords:

Streptokinase, Recombinant protein, Affinity chromatography, Hydrophobic gel purification, Ammonium sulphate

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.

Author Biography

Nastaran Rahimi1 , Mohammad AlinezhadChamazketi2, Amir Yaghoubi Nezhad3, Forough Talaeizadeh4

1Research assistant in the Islamic Azad University Pharmaceutical Sciences Branch, Tehran, Iran, 2 Lecturer in Department of Biotechnology, Malek Ashtar University of Technology (MUT), Tehran, Iran, 3Department of Cellular and Molecular Biology, Faculty of Sciences, Higher Education Institute of Rab-Rashid, Tabriz, Iran, 4Department of Educational Science and Psychology, Islamic Azad University Marvdasht Branch, Marvdasht, Iran

Published

2020-04-29

How to Cite

Nastaran Rahimi1 , Mohammad AlinezhadChamazketi2, Amir Yaghoubi Nezhad3, Forough Talaeizadeh4. (2020). Optimization of Streptokinase Mutant Protein Purification Method Using Affinity Chromatography Technique. Indian Journal of Forensic Medicine & Toxicology, 14(2), 2369-2373. https://doi.org/10.37506/ijfmt.v14i2.3383