Fe3O4@Al2O3 magnetic core-shell microspheres for rapid and highly specific capture of phosphopeptides with mass spectrometry analysis.
Fe3O4@Al2O3 magnetic core-shell microspheres for rapid and highly specific capture of phosphopeptides with mass spectrometry analysis.
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DOI：
10.1016/j.chroma.2007.09.062
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发表时间：
2007-11
期刊：
影响因子：
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通讯作者：
Yan Li;Yingchao Liu;Jia Tang;huaqing lin;N. Yao;Xizhong Shen;Chunhui Deng;Pengyuan Yang;
中科院分区：
文献类型：
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作者：
Yan Li;Yingchao Liu;Jia Tang;huaqing lin;N. Yao;Xizhong Shen;Chunhui Deng;Pengyuan Yang;
在许多蛋白质组学应用中，从复杂生物样品中选择性检测磷脂肽是一项具有挑战性且高度相关的任务。本研究开发了一种基于Fe3O4@Al2O3magnetic核壳微球与磷酸肽强相互作用的新型磷酸肽富集方法。Fe3O4@Al2O3magnetic核壳微球具有明确的核壳结构，不仅具有氧化铝外壳，使其具有对磷酸肽的高捕获能力，而且还具有磁性，可以通过定位外部磁场轻松隔离。制备的Fe3O4@Al2O3magnetic核壳微球已成功应用于标准磷酸化蛋白β-酪蛋白和卵清蛋白的胰蛋白酶消化中磷酸肽的富集。通过分析β-酪蛋白与牛血清白蛋白摩尔比为1:50的消化混合物中的磷酸肽，以及酪蛋白与5种蛋白质混合物的胰蛋白酶消化产物，证明了该方法具有良好的选择性。结果还证明Fe3O4@Al2O3magnetic核壳微球对Fe3+固定化磁性二氧化硅微球、商用Fe3+ -IMAC（固定化金属亲和层析）树脂和tio2微珠具有更强的选择性。最后，利用al2o3包被fe3o4微球，成功富集了大鼠肝提取物消化产物中的磷酸肽。这些结果表明Fe3O4@Al2O3magnetic核壳微球是一种快速、选择性分离和富集磷酸肽的良好材料。
Selective detection of phosphopeptides from complex biological samples is a challenging and highly relevant task in many proteomics applications. In this study, a novel phosphopeptide enrichment approach based on the strong interaction of Fe3O4@Al2O3magnetic core–shell microspheres with phosphopeptides has been developed. With a well-defined core–shell structure, the Fe3O4@Al2O3magnetic core–shell microspheres not only have a shell of aluminum oxide, giving them a high-trapping capacity for the phosphopeptides, but also have magnetic property that enables easy isolation by positioning an external magnetic field. The prepared Fe3O4@Al2O3magnetic core–shell microspheres have been successfully applied to the enrichment of phosphopeptides from the tryptic digest of standard phosphoproteins β-casein and ovalbumin. The excellent selectivity of this approach was demonstrated by analyzing phosphopeptides in the digest mixture of β-casein and bovine serum albumin with molar ratio of 1:50 as well as tryptic digest product of casein and five protein mixtures. The results also proved a stronger selective ability of Fe3O4@Al2O3magnetic core–shell microspheres over Fe3+-immobilized magnetic silica microspheres, commercial Fe3+–IMAC (immobilized metal affinity chromatography) resin, and TiO2beads. Finally, the Al2O3coated Fe3O4microspheres were successfully utilized for enrichment of phosphopeptides from digestion products of rat liver extract. These results show that Fe3O4@Al2O3magnetic core–shell microspheres are very good materials for rapid and selective separation and enrichment of phosphopeptides.