Preparation of Fe3O4@ZrO2 Core-Shell Microspheres as Affinity Probes for Selective Enrichment and Direct Determination of Phosphopeptides Using Matrix-Assisted Laser D

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• 作者：Yan Li ; Taohua Leng ; Huaqing Lin ; Chunhui Deng ; Xiuqing Xu ; Ning Yao ; Pengyuan Yang ; Xiangmin Zhang
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：November 2007
• 年：2007
• 卷：6
• 期：11
• 页码：4498 - 4510
• 全文大小：781K
• 年卷期：v.6,no.11(November 2007)
• ISSN：1535-3907

文摘

Fe₃O₄@ZrO₂ microspheres with well-defined core-shell structure were prepared and applied for the highlyselective enrichment of phosphopeptides from trypticdigest product of proteins. To successfully coat iron oxidemicrospheres with uniform zirconia shell, magnetic Fe₃O₄microspheres were first synthesized via a solvothermalreaction, followed by being coated with a thin layer ofcarbon by polymerization and carbonization of glucosethrough hydrothermal reaction. Finally, with the use ofthe Fe₃O₄@C microspheres as templates, zirconium isopropoxide was prehydrolyzed and absorbed onto themicrospheres and eventually converted into zirconia bycalcinations. The as-prepared Fe₃O₄@ZrO₂ core-shellmicrospheres were used as affinity probes to selectivelyconcentrate phosphopeptides from tryptic digest of-casein, casein, and five protein mixtures to exemplifytheir selective enrichment ability of phosphopeptides fromcomplex protein samples. In only 0.5 min, phosphopeptides sufficient for characterization by MALDI-MS couldbe enriched by the Fe₃O₄@ZrO₂ microspheres. The resultsdemonstrate that Fe₃O₄@ZrO₂ microspheres have theexcellent selective enrichment capacity for phosphopeptides from complex samples. The performance of theFe₃O₄@ZrO₂ microspheres was further compared withcommercial IMAC beads for the enrichment of peptidesoriginating from tryptic digestion of -casein and bovineserum albumin (BSA) with a molar ratio of 1:50, and theresults proved a stronger selective ability of Fe₃O₄@ZrO₂microspheres over IMAC beads. Finally, the Fe₃O₄@ZrO₂microspheres were successfully utilized for enrichmentof phosphopeptides from human blood serum withoutany other purification procedures.