Synthetics of NiFe2O4 nanoparticles for recombinant His-tag protein purification

Document Type : Reasearch Paper

Authors

• S. Imani ¹
• A.M. Zand ¹
• M. Saadati ²
• H. Honnari ¹
• B. Maddah ³

¹ Department and Research Center of Biology, IHU, Tehran, Iran.

² Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.

³ Department of chemistry, IHU, Tehran, Iran.

10.7508/ijnd.2011.02.006

Abstract

Recombinant protein purification is a kind of sensitive and expensive method in genetics engineering. Genetic manipulation leads to the expression of various proteins; it should be isolated with high purity finally. Differed methods for protein purification are categorized, based on cast, quality, Easy work and side effect of protein. In this article, we are investigating his His-tag protein purification by magnetic nanoparticles. NiFe₂O₄ nanoparticles were synthesized by Co-precipitation method, than was dissolved in 0.05 NaCl solutions. Tube containing of nanoparticles and buffer was located in magnetic field. Nanoparticles were separation by the three-stage washed. According to the protocols, nanoparticles located in his-tag protein and the end protein were predicated, and analyzed by the Electrophoreses.   The results of gel showed can be extracted that the proteins form a weak that the results of gel showed that proteins isolated are poor by this method. By supplementary study, we get new age or tree-age nanoparticles, that Ni is the surface of N.P. This age of nanoparticles is fit to protein purification. Nano-particle synthesis in this article, are created a two-dimensional grid of nickel and iron oxide nanoparticles that this structure is not suitable for purification of His-tag protein. So the magnetic nanoparticles have a three-dimensional structure of the nickel nanoparticles on the surface of the exposed histidine to be enough space to connect and link. It is hoped in future studies with these types of nanoparticles synthesis to achieved His-protein isolation kit.

Keywords

• MNP
• Recombinant protein
• Protein purification

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