Purification of Histidine-tagged Fusion Proteins Based on Fe3O4@SiO2/Ni-NTA Magnetic Spheres

doi:10.3969/j.issn.0251-0790.2012.02.016

Abstract

Abstract: The development of reliable and efficient methods to separate proteins of interest from a biological source is still a challenging work. Currently, using the His-tagged fusion proteins to separate and purifiy proteins has been the most widely used technology. This study deals with the synthesis of Ni-nitrilotriacetic acid(Ni-NTA) modified Fe₃O₄@SiO₂ magnetic spheres, which can act as a general agent to separate and purify Histidine-tagged fusion proteins. The spheres have a core composed of carbon particles and a shell composed of Fe₃O₄ having a uniform size of ca.402 nm, which provides the spheres with excellent magnetic responsivity and dispersity. The recombinant proteins that had been engineered to have six consecutive histidine residues(6×His) were separated by means of Fe₃O₄@SiO₂/Ni-NTA magnetic spheres. The results show that 10 mg of Fe₃O₄@SiO₂/Ni-NTA spheres are able to purify about 1 mg of 6×His-tagged proteins from 10 mL crude E. coli. lysate. Owing to the high separation capacity, Fe₃O₄@SiO₂/Ni-NTA spheres can be used to separate 6×His-tagged proteins with low-concentrations.

Key words: Magnetic sphere, 6×His-tagged protein, Affinity Purification, Ni-nitrilotriacetic acid

CLC Number:

O629

TrendMD:

WANG Wen-Jia, GUO Xiao-Lin, WEI An-Hui, FU Chang-Hao, HAN Zhen-Guo. Purification of Histidine-tagged Fusion Proteins Based on Fe₃O₄@SiO₂/Ni-NTA Magnetic Spheres[J]. Chem. J. Chinese Universities, 2012, 33(02): 303.

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Purification of Histidine-tagged Fusion Proteins Based on Fe₃O₄@SiO₂/Ni-NTA Magnetic Spheres

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