Highly selective and rapid enrichment of phosphorylated peptides using gallium oxide‐coated magnetic microspheres for MALDI‐TOF‐MS and nano‐LC‐ESI‐MS/MS/MS analysis
Highly selective and rapid enrichment of phosphorylated peptides using gallium oxide‐coated magnetic microspheres for MALDI‐TOF‐MS and nano‐LC‐ESI‐MS/MS/MS analysis
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DOI：
10.1002/pmic.200700454
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发表时间：
2008
期刊：
影响因子：
3.4
通讯作者：
Yan Li;huaqing lin;Chunhui Deng;Pengyuan Yang;Xiangmin Zhang
中科院分区：
文献类型：
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作者：
Yan Li;huaqing lin;Chunhui Deng;Pengyuan Yang;Xiangmin Zhang
在这项工作中，我们提出，据我们所知，第一次演示的实用性的氧化铁磁性微球涂有氧化镓的高选择性富集磷酸肽之前，质谱分析。我们制备的这些微球不仅具有氧化镓的外壳，使它们对磷酸肽具有高捕获能力，而且它们的磁性可以通过定位外部磁场轻松分离。使用包括β-酪蛋白、卵清蛋白、酪蛋白以及五种蛋白质混合物的磷蛋白的胰蛋白酶消化产物作为样品来验证该方法的可行性。在非常短的时间内（仅0.5分钟），足以通过MALDI-TOF-MS表征的磷酸肽被Ga 2 O3-包覆的Fe 3 O 4微球选择性富集。进一步比较了Ga 2 O3包覆的Fe 3 O 4微球与固定化Fe 3+的磁性二氧化硅微球、市售Fe 3 +-IMAC树脂和TiO 2微球对胰蛋白酶消化的β-酪蛋白和BSA（摩尔比为1：50）产生的肽的富集性能，结果证明Ga 2 O3包覆的Fe 3 O 4微球比其他材料具有更强的选择能力。最后，Ga 2 O3包覆的Fe 3 O 4微球被成功地用于从大鼠肝脏提取物的消化产物中富集磷酸肽。结果表明，Ga 2 O3包覆Fe 3 O 4微球是一种有效的选择性分离富集磷酸肽的材料。
In this work, we present, to our knowledge, the first demonstration of the utility of iron oxide magnetic microspheres coated with gallium oxide for the highly selective enrichment of phosphopeptide prior to mass spectrometric analysis. These microspheres that we prepared not only have a shell of gallium oxide, giving them a high‐trapping capacity for the phosphopeptides, but also their magnetic property enables easy isolation by positioning an external magnetic field. Tryptic digest products of phosphoproteins including β‐casein, ovalbumin, casein, as well as five protein mixtures were used as the samples to exemplify the feasibility of this approach. In very short time (only 0.5 min), phosphopeptides sufficient for characterization by MALDI‐TOF‐MS were selectively enriched by the Ga2O3‐coated Fe3O4 microspheres. The performance of the Ga2O3‐coated Fe3O4 microspheres were further compared with Fe3+‐immobilized magnetic silica microspheres, commercial Fe3+‐IMAC resin, and TiO2 beads for enrichment of peptides originating from tryptic digestion of β‐casein and BSA with a molar ratio of 1:50, and the results proved a stronger selective ability of Ga2O3‐coated Fe3O4 microspheres over the other materials. Finally, the Ga2O3‐coated Fe3O4 microspheres were successfully utilized for enrichment of phosphopeptides from digestion products of rat liver extract. All results show that Ga2O3‐coated Fe3O4 microsphere is an effective material for selective isolation and concentration of phosphopeptides.