Fe₃O₄/PVIM-Ni²⁺ magnetic composite microspheres for highly specific separation of histidine-rich proteins.
Fe₃O₄/PVIM-Ni²⁺ magnetic composite microspheres for highly specific separation of histidine-rich proteins.
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DOI：
10.1021/am501626t
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发表时间：
2014-05
影响因子：
9.5
通讯作者：
Y. Zhang;D. Li;Meng Yu;Wanfu Ma;Jia Guo;Changchun Wang
中科院分区：
文献类型：
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作者：
Y. Zhang;D. Li;Meng Yu;Wanfu Ma;Jia Guo;Changchun Wang
结合固定化金属离子亲和层析和磁性微球的优点，被认为是一种快速、方便地分离组氨酸标记蛋白质的理想途径，但很少有报道涉及天然富含组氨酸的蛋白质。采用回流-沉淀聚合法，制备了由高磁响应性磁性超微粒(Fe3O4)核和Ni(2+)负载的交联咪唑(PVIM)壳组成的磁性微球。所制备的Fe3O4/PVIM-Ni(2+)微球具有均匀的花状结构、高的磁响应性、丰富的结合部位和非常容易的合成工艺。该微球具有纯的PVIM-Ni(2+)界面和较高的Ni(2+)负载量，在His标记的重组蛋白的固定化和分离中具有显著的选择性、良好的灵敏度、大的富集量和高的可回收性。更有趣的是，研究发现，由于暴露的组氨酸残基，Fe3O4/PVIM-Ni(2+)微球对从复杂的胎牛血清中去除天然富含组氨酸的牛血清白蛋白(BSA)也表现出了良好的性能。考虑到它们的多重优点，这种新型的Fe3O4/PVIM-Ni(2+)纳米材料在富含低丰度的组氨酸标记的蛋白质或去除富含组氨酸的天然蛋白质进行蛋白质组学分析方面具有巨大的潜力。
Integration of the advantages of immobilized metal-ion affinity chromatography (IMAC) and magnetic microspheres is considered as an ideal pathway for quick and convenient separation of his-tagged proteins, but rare reports concern the natural histidine-rich proteins. In this article, a novel route was presented to fabricate magnetic microspheres composed of a high-magnetic-response magnetic supraparticle (Fe3O4) core and a Ni(2+)-immobilized cross-linked polyvinyl imidazole (PVIM) shell via reflux-precipitation polymerization. The unique as-prepared Fe3O4/PVIM-Ni(2+) microspheres possessed uniform flower-like structure, high magnetic responsiveness, abundant binding sites, and very easy synthesis process. Taking advantage of the pure PVIM-Ni(2+) interface and high Ni(2+) loading amount, the microspheres exhibited remarkable selectivity, excellent sensitivity, large enrichment capacity, and high recyclability in immobilization and separation of his-tagged recombinant proteins. More interestingly, it was found that the Fe3O4/PVIM-Ni(2+) microspheres also showed excellent performance for removal of the natural histidine-rich bovine serum albumin (BSA) from the complex real sample of fetal bovine serum due to the exposed histidine residues. Considering their multiple merits, this new type of Fe3O4/PVIM-Ni(2+) nanomaterial displays great potential in enriching low-abundant his-tagged proteins or removing high-abundant histidine-rich natural proteins for proteomic analysis.