血清蛋白质组学研究中乙腈沉淀法去除清蛋白的实验研究
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Establishment of acetonitrile precipitation method for removing albumin in serum proteomics research
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    摘要:

    目的 建立去除人血清清蛋白的乙腈沉淀法并研究其去除效果。 方法 血清经除脂后按血清∶水∶乙腈=1∶2∶4.5(v/v/v)比例沉淀分离高丰度清蛋白,用SDS-PAGE验证去除效果和重复性。分别对4名体检健康者和非小细胞肺癌患者去除清蛋白前后的血清凝胶电泳图像进行分析。 结果 经本法去除后血清清蛋白平均浓度由(50.65±2.82) g/L下降为(2.50±0.71) g/L,占血清总蛋白比例由65.1%下降为33.3%,呈显著降低,差异有统计学意义(P<0.001)。与等量上样原血清电泳结果相比,相对分子量(Mr)在50 000~80 000间的蛋白质被明显去除,而被其掩盖的低丰度蛋白质得以显现,蛋白质条带明显变窄。 结论 乙腈沉淀法提供了一种经济、简便,且重复性较好的人血清清蛋白去除方法,为血清蛋白质组学研究提供了技术支持。

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    Objective To establish and validate a simple and economical acetonitrile precipitation method to deplete albumin from serum and investigate its effectiveness. Methods The albumin was precipitated by mixing delipidated serum, water, acetonitrile at ratio of 1∶2∶4.5 (v/v/v). The removal effects and repeatability were analyzed by onedimensional sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). The gel images of serum samples from 4 healthy controls and 4 patients with nonsmallcell lung carcinoma were analyzed before and after treatment. Results The results showed that the average concentration of albumin in the sera was decreased from (50.65 ± 2.82) g/L to (2.50 ± 0.71) g/L. The percentage of albumin in the total protein of serum was decreased from 65.1% to 33.3%, and significantly statistical differences were found (P<0.01). Compared with the gel images of the original serum with same volume, the band of protein with molecular weight between 50-80 kDa was removed. The band narrowed significantly and low abundance proteins could be seen. Conclusions The acetonitriledepletion strategy may offer an economical, simple and reproducible method to remove albumin from human serum, which provided a technical support for further study of serum proteomics.

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朱慧,李克,翟云霞.血清蛋白质组学研究中乙腈沉淀法去除清蛋白的实验研究[J].临床检验杂志,2011,29(1):16-18

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  • 收稿日期:2010-04-11
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