Depletion of the high-abundance plasma proteins
Depletion of the high-abundance plasma proteins
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DOI：
10.1007/s00726-004-0141-1
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发表时间：
2004-12-01
期刊：
影响因子：
3.5
通讯作者：
Langen, H
中科院分区：
文献类型：
--
作者：
Fountoulakis, M;Juranville, JF;Langen, H
体液，如血浆和尿液，相对容易获得，并且可用于通过应用新技术（如蛋白质组学）检测新的诊断标志物。然而，在血浆中，几种高丰度蛋白质占主导地位，并抑制低丰度蛋白质的信号，然后通过二维凝胶或色谱法变得无法检测。因此，大量蛋白质的消耗是检测低丰度组分的先决条件。我们在蓝色基质和蛋白G上应用亲和层析，并去除最丰富的人血浆蛋白、白蛋白和免疫球蛋白链。血浆蛋白，白蛋白和免疫球蛋白消耗之前，以及从两个色谱步骤的洗脱液进行了分析，通过二维电泳和蛋白质进行了鉴定，通过MALDI-TOF-MS。分析导致在未处理的血浆中的83个不同的基因产物的识别。去除高丰度蛋白质导致新蛋白质信号的可视化。在两个亲和步骤的第二步中，主要检测到白蛋白和免疫球蛋白斑点，但也检测到代表其他几种丰富血浆蛋白的斑点。该方法易于执行，并且作为通过应用蛋白质组学技术检测体液中的诊断标志物的第一步是有用的。
Body fluids, like plasma and urine, are comparatively easy to obtain and are useful for the detection of novel diagnostic markers by applying new technologies, like proteomics. However, in plasma, several high-abundance proteins are dominant and repress the signals of the lower-abundance proteins, which then become undetectable either by two-dimensional gels or chromatography. Therefore, depletion of the abundant proteins is a prerequisite for the detection of the low-abundance components. We applied affinity chromatography on blue matrix and Protein G and removed the most abundant human plasma proteins, albumin and the immunoglobulin chains. The plasma proteins, prior to albumin and immunoglobulin depletion, as well the eluates from the two chromatography steps were analyzed by two-dimensional electrophoresis and the proteins were identified by MALDI-TOF-MS. The analysis resulted in the identification of 83 different gene products in the untreated plasma. Removal of the high-abundance proteins resulted in the visualization of new protein signals. In the eluate of the two affinity steps, mostly albumin and immunoglobulin spots were detected but also spots representing several other abundant plasma proteins. The methodology is easy to perform and is useful as a first step in the detection of diagnostic markers in body fluids by applying proteomics technologies.