基于Fe_3O_4@SiO_2/Ni-NTA磁性微球的His-tag融合蛋白纯化体系的建立  

Purification of Histidine-tagged Fusion Proteins Based on Fe_3O_4@SiO_2/Ni-NTA Magnetic Spheres

作　　者：王文加[1] 郭晓林[2] 韦安慧[1] 付常皓[1] 韩振国[3] 

机构地区：[1]吉林大学药学院 [2]吉林大学第一医院,长春130021 [3]吉林大学中日联谊医院,长春130031

出　　处：《高等学校化学学报》2012年第2期303-307,共5页Chemical Journal of Chinese Universities

摘　　要：合成了表面共价结合Ni-氨基三乙酸(Ni-NTA)基团的Fe3O4@SiO2微球,这种磁性微球可用于分离含有His-tag标签的融合蛋白.微球中心由尺寸约402 nm的Fe3O4微粒组成,赋予了微球极好的磁性分离和离心分离的特性.应用Fe3O4@SiO2/Ni-NTA磁性微球对含有6×His-tag(6聚组氨酸)标签的蛋白进行了分离纯化,结果表明,10 mg Fe3O4@SiO2/Ni-NTA微球能够从10 mL重组蛋白裂解液中纯化出约1 mg带有6×His-tag标签的融合蛋白.微球的高效分离效果使其能够用于含量较低的带有6×His-tag标签蛋白的分离纯化.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.

关 键 词：磁性微球 6×His-tag蛋白 亲和纯化 Ni-氨基三乙酸 

分 类 号：O629[理学—有机化学]