Quality control of serum albumin, depletion for proteomic analysis
Quality control of serum albumin, depletion for proteomic analysis
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DOI：
10.1373/clinchem.2007.091736
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发表时间：
2007-11-01
影响因子：
9.3
通讯作者：
Suffredini, Anthony F.
中科院分区：
文献类型：
--
作者：
Seam, Nitin;Gonzales, Denise A.;Suffredini, Anthony F.
背景：预分离技术，如血清白蛋白去除是蛋白质组学分析的有用前体，但如果去除不可重复，它们可能会引入分析前偏差。我们检查了白蛋白免疫消耗的可重复性，并描述了一种质量控制方法。方法：用IgY免疫亲和自旋柱对合并血清中白蛋白的损耗进行评估，每柱21次。我们在Beckman LX 20分析仪上使用免疫比浊法测定白蛋白耗尽后的白蛋白浓度，并使用SELDI-TOF质谱法评估白蛋白耗尽样品的质谱。结果：白蛋白洗脱效率存在显著的运行间差异，多次使用后效率系统性下降。在前17趟井中，有15趟井的平均衰竭效率为95%
Background: Prefractionation techniques such as serum albumin depletion are useful precursors to proteomic analysis, but they may introduce preanalytical bias if the depletion is not reproducible. We examined the reproducibility of albumin immunodepletion and describe a method of QC for this process.'Methods: Depletion of albumin from pooled serum, performed using IgY immunoaffinity spin columns, was assessed for 21 runs on each of 4 columns. We measured albumin concentrations, after albumin depletion, by use of an immunoturbidimetric assay on the Beckman LX 20 analyzer and assessed mass spectra of albumin-depleted samples by use of SELDI-TOF mass spectrometry.Results: There was substantial run-to-run variation in efficiency of albumin depletion, with systematic decline in efficiency after multiple uses of the columns. Mean depletion efficiency was >95% for 15 of the 1st 17 runs and