Analytical and Bioanalytical Chemistry ( IF 3.8 ) Pub Date : 2021-09-07 , DOI: 10.1007/s00216-021-03637-5 Liang Ma 1 , Yindi Zhu 1 , Xueming Chen 1 , Raohao Fang 1 , Yuru Chen 1 , Xia Xu 1, 2 , Guozheng Huang 1 , Zi Liu 1 , Xiang Liu 1
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Nano-magnetite with superparamagnetism could be coated by some organic compounds or by nano Au or Pt via surface modifications with multi-step reactions for the applications of isolating histidine-tagged (His-tagged) proteins. Introducing active sites of binding histidine onto the surface of nano-magnetite was the ultimate task. However, multi-step treatments might result in departure of the coatings from the surface of the nano-magnetite, which led to loss of active sites. In this work, we reported a convenient and efficient way of treating nano-magnetites and applied them in isolating His-tagged proteins. Carboxylates were introduced on the surface of home-made nano-magnetite directly via ultrasonic mixing with sodium bitartrate rather than complicated surface modifications, which was proved by thermogravimetric analyses. Ni2+ was, therefore, caught by the carboxylates of the coating via the coordinate interaction, demonstrated by X-ray photoelectron spectra. The coated magnetic nanoparticles with the bonded Ni2+ were successfully employed to selectively bind and separate recombinant His-tagged proteins directly from the mixture of Escherichia coli cell lysate, and showed wonderful affinity for His-tagged proteins with the saturated adsorption amount being 556 mg g−1. Additionally, such functionalized nano-magnetite manifested the excellent recyclability in isolating His-tagged proteins.
Graphical abstract
中文翻译：
一种用于分离大肠杆菌中表达的组氨酸标签蛋白的新型镍改性纳米磁铁
具有超顺磁性的纳米磁铁矿可以被一些有机化合物或纳米金或铂通过多步反应的表面修饰来涂覆，用于分离组氨酸标记（His-tagged）的蛋白质。将结合组氨酸的活性位点引入纳米磁铁矿表面是最终任务。然而，多步骤处理可能会导致涂层从纳米磁铁矿表面脱离，从而导致活性位点的损失。在这项工作中，我们报道了一种处理纳米磁铁矿的方便有效的方法，并将其应用于分离 His 标签蛋白。羧酸盐是通过与酒石酸氢钠的超声混合直接引入自制纳米磁铁矿表面的，而不是复杂的表面改性，热重分析证明了这一点。你因此，通过 X 射线光电子能谱证明，2 +通过配位相互作用被涂层的羧酸盐捕获。结合Ni 2+的包覆磁性纳米粒子成功地直接从大肠杆菌细胞裂解液中选择性结合和分离重组His-tagged蛋白，对His-tagged蛋白表现出极好的亲和力，饱和吸附量为556 mg g -1。此外，这种功能化的纳米磁铁矿在分离带组氨酸标签的蛋白质方面表现出优异的可回收性。
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