Limitation of immunoaffinity column for the removal of abundant proteins from plasma in quantitative plasma proteomics
Limitation of immunoaffinity column for the removal of abundant proteins from plasma in quantitative plasma proteomics
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DOI：
10.1002/bmc.1139
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发表时间：
2009-05-01
影响因子：
1.8
通讯作者：
Imai, Kazuhiro
中科院分区：
文献类型：
--
作者：
Ichibangase, Tomoko;Moriya, Kyoji;Imai, Kazuhiro
在血浆蛋白质组学中，在蛋白质组学分析之前，必须通过特定的制备技术，如免疫亲和捕获，制备不含高丰度蛋白质（包括白蛋白）的蛋白质样品。然而，我们的初步实验表明，随着使用后的免疫亲和柱的能力的功能变化。因此，在本研究中，为了评价免疫亲和柱从血浆中去除丰富蛋白的能力的变化，使用荧光衍生-液相色谱-串联质谱法结合IgY柱进行血浆蛋白质组分析，以长期测试亲和柱的重现性。白蛋白的特异性吸附随色谱柱使用次数的增加而降低。此外，它被证明是疏水性的高分子量化合物的血浆吸附到柱材料表面上的功能从特异性的免疫亲和吸附到疏水相互作用的变化。这些结果表明，在定量血浆蛋白质组学研究中，重要的是要记住的风险，不仅非选择性损失，而且在免疫亲和柱的吸附能力的变化。版权所有（C）2008约翰威利父子有限公司
In plasma proteomics, before a proteome analysis, it is essential to prepare protein samples without high-abundance proteins, including albumin, via specific preparation techniques, such as immunoaffinity capture. However, our preliminary experiments suggested that functional changes with use after the ability of the immunoaffinity column. Thus, in this study, to evaluate the changes of the removal ability of abundant proteins from plasma by the immunoaffinity column, plasma proteome analysis was performed for the long-term test for the reproducibility of the affinity column using the fluorogenic derivatization-liquid chromatography-tandem mass spectrometry method combined with an IgY column. The specific adsorption for albumin decreased with an increase in the number of the column usage before its expiration date. Moreover, it was demonstrated that hydrophobic high molecular weight compounds in plasma adsorbed onto the column materials surface contributed to the functional changes from specific immunoaffinity adsorption into hydrophobic interaction. These results suggested that, in quantitative plasma proteomics studies, it is important to keep in mind the risk of not only the nonselective loss but also the changes in the adsorption ability of the immunoafinity column. Copyright (C) 2008 John Wiley & Sons, Ltd.