Fe3O4/Au Core/Shell Nanoparticles Modified with Ni2+-Nitrilotriacetic Acid Specific to Histidine-Tagged Proteins
Fe3O4/Au Core/Shell Nanoparticles Modified with Ni2+-Nitrilotriacetic Acid Specific to Histidine-Tagged Proteins
复制标题
用 Ni2-次氮基三乙酸修饰的 Fe3O4/Au 核/壳纳米颗粒，对组氨酸标记蛋白具有特异性
DOI：
10.1021/jp910753f
复制
发表时间：
2010-03-25
影响因子：
3.7
通讯作者：
Hao, Jian
中科院分区：
文献类型：
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作者：
Xie, Hai-Yan;Zhen, Rui;Hao, Jian
以聚乙烯亚胺(PEI)为连接剂，成功制备了具有良好单分散性和较强磁性的Fe3O4/Au核/壳纳米粒子。完整的金壳使这些纳米颗粒易于修饰和生物功能化，以满足不同的生物检测和生物传感目的。在此基础上，用巯基丙酸对其进行表面修饰，然后对其进行偶联氨基三乙酸(NTA)，然后再对Ni2+进行络合。将得到的生物功能化Fe3O4/Au-NTA-Ni2+复合纳米颗粒用于直接从裂解细胞混合物中富集和分离组氨酸标记的麦芽糖结合蛋白(MBP)。结果表明，Fe3O4/Au-NTA-Ni2+可用于快速、高效、特异地富集和分离His-Tag融合蛋白。Fe3O4/Au-NTA-Ni2+纳米粒子的富集率明显高于金属络合亲和层析(MCAC)。目前的方法与简易的SDS-PAGE联用，检测下限可低于5.5×10(-8)M。由于操作简单，分离效率高，可以进一步开发多种双功能甚至多功能纳米材料用于生物应用。
Well-defined Fe3O4/Au core/shell nanoparticles were successfully prepared with polyethyleneimine (PEI) as a linker, which are of good monodispersity and strong magnetism. The intact gold shell made these nanoparticles easily modified and biofunctionalized for different biodetection and biosensing purposes. Hereby, they were surface modified with mercaptopropionic acid, followed by conjugating nitrilotriacetic acid (NTA) and subsequently chelating Ni2+. The resulting biofunctionalized Fe3O4/Au-NTA-Ni2+ composite nanoparticles were used to enrich and separate the histidine-tagged (His-Tag) maltose-binding protein (MBP) directly from the mixture of lysed cells. It has been found that Fe3O4/Au-NTA-Ni2+ can be used for rapid, efficient, and specific enrichment and separation of His-Tag fusion proteins. The enrichment efficiency of the Fe3O4/Au-NTA-Ni2+ nanoparticles is significantly higher that that of metal-chelate affinity chromatography (MCAC). The detection limit of the current method coupled with facile SDS-PAGE can be lower than 5.5 x 10(-8)M. Due to the ease of operation and good efficiency of separation, diverse bifunctional and even multifunctional nanomaterials can be further developed for biological applications.