巯基功能化磁性微球的构建及其对蛋白质的亲和分离  

Construction of thiol-functional magnetic microspheres and affinity separation of protein

作　　者：刘珊珊[1] 邹雪艳[2] 郭静玉[1] 刘锦[1] 陈丹云[1] 

机构地区：[1]河南大学化学化工学院,河南开封475004 [2]河南大学特种功能材料重点实验室,河南开封475004

出　　处：《化学研究》2015年第5期534-539,共6页Chemical Research

基　　金：国家自然科学基金(21271062);河南省教育厅科学技术重点研究项目(14B150003)

摘　　要：通过水热法一步合成了纳米Fe3O4微球,并在甲苯中用巯基丙基三甲氧基硅烷(MPS)对其进行表面修饰得到了Fe3O4-SH微球,通过DTNB法测得微球表面巯基含量为333.54μg/mg.该纳米微球可以吸附溶液中的Ni2+,从而形成Fe3O4-SH-Ni2+复合材料.以此复合材料为载体,可以将以组氨酸为标签的(His-tagged)融合蛋白直接从细胞裂解液中进行提纯,并在外加磁场的作用下实现对目标蛋白的快速分离,其对His-tagged TRX蛋白的分离能力为20.6μg/mg,特别适合于对以组氨酸为标签蛋白的分离纯化.Ferriferrous oxide magnetic microspheres （MSs） were prepared via solvothermal route .The so‐obtained MSs were modified by （3‐mercapto‐propyl）trimethoxysilane （MPS） in mythylbenzene to give birth of the ferriferrous oxide with thiol group （Fe3 O4‐SH ） micro‐spheres ,which can absorb Ni2＋ to form Fe3 O4‐SH‐Ni2＋ .The amount of thiol group of Fe3 O4‐SH was tested to be 333 5.4 μg/mg by means of DTNB method .After chelating Ni2＋ ions ,Fe3 SH‐Ni2＋ MSs were used to enrich and purify histidine‐tagged （His‐tagged） proteins directly from the mixture of lysed cells without pretreatment .Moreover ,the prepared MSs can sepa‐rate the target protein quickly in the magnetic field .It has been found that Fe3 O4‐SH‐Ni2＋ MSs present negligible nonspecific protein adsorption and high protein binding activity with the sat‐uration capacity being 20 6.μg/mg and they are especially suitable for rapid purification of His‐tagged proteins .

关 键 词：四氧化三铁 巯基 组氨酸 蛋白质分离 亲和 

分 类 号：O61[理学—无机化学]