Lee Jiho, Lee Soo Youn, Park So Hyun, Lee Hye Sun, Lee Jin Hyung, Jeong Bong-Yong, Park Sang-Eon, Chang Jeong Ho
Bio-IT Convergence Center, Korea Institute of Ceramic Engineering and Technology, Seoul 153-801, Korea.
J Mater Chem B. 2013 Feb 7;1(5):610-616. doi: 10.1039/c2tb00232a. Epub 2012 Nov 23.
This work reports a simple and facile method to prepare novel magnetic mesoporous silica (MMS) materials with high magnetic strength for the convenient and high throughput detection of histidine-tagged enzymes with Ni-doped surfaces. These materials are designed by the incorporation of high-abundance and homogeneously dispersed iron nanoparticles within the mesopores by thermal hydrogen reduction after the incorporation of ferrous ions and demonstrated the selective enrichment and high-throughput recognition of His-tagged enzymes with multi-point anchoring by selective conjugation between the His-tag and Ni ions. Selective His-tagged enzyme enrichment efficiency was compared with nickel-based MMS materials, such as Ni-MMS and Ni-MMS, and nickel ion doped silica-coated magnetic nanoparticles (Ni-MNPs). The efficiency was calculated to be 100 ± 1.93%, 70.94 ± 1.95%, and 37.03 ± 5.93% for Ni-MMS, Ni-MMS, and Ni-MNPs, respectively. This method enables a high-throughput and advanced systematic approach for the separation and immobilization of proteins which cover a broad spectrum of polyhistidine-tagged proteins.
本工作报道了一种简单易行的方法，用于制备具有高磁强度的新型磁性介孔二氧化硅（MMS）材料，以便于对表面掺杂镍的组氨酸标签酶进行便捷且高通量的检测。这些材料是通过在掺入亚铁离子后，经热氢还原将高丰度且均匀分散的铁纳米颗粒掺入介孔中而设计的，并通过组氨酸标签与镍离子之间的选择性共轭，证明了对组氨酸标签酶的多点锚定选择性富集和高通量识别。将选择性组氨酸标签酶富集效率与镍基MMS材料（如Ni-MMS和Ni-MMS）以及镍离子掺杂的二氧化硅包覆磁性纳米颗粒（Ni-MNPs）进行了比较。Ni-MMS、Ni-MMS和Ni-MNPs的效率分别计算为100±1.93%、70.94±1.95%和37.03±5.93%。该方法为涵盖广泛多聚组氨酸标签蛋白的蛋白质分离和固定提供了一种高通量且先进的系统方法。
