脑脊液双向电泳中去除高丰度蛋白技术的研究

High-abundant protein depletion method for cerebrospinal fluid in 2-DE analysis

目的建立一种去除脑脊液高丰度蛋白的方法并进行双向电泳分析评价。方法采用超滤法浓缩脑脊液样品至适当体积,用商业化血清白蛋白和免疫球蛋白G(IgG)去除试剂盒处理,得到含低丰度蛋白的脑脊液,再用丙酮沉淀法除盐并进行二维凝胶电泳(2-DE)分析和2-DE Western blot鉴定。结果此法可有效去除脑脊液中的白蛋白和IgG,2-DE图谱分析显示,去除高丰度蛋白后脑脊液低丰度蛋白富集明显,与2-DE Western blot结果对比,四连接素的三个不同片段去除高丰度蛋白后可全部显现。结论本研究建立的脑脊液高丰度蛋白去除方法简便易行,有利于双向电泳中低丰度蛋白的检出。

Objective To establish and evaluate a method of depleting high-abundant proteins for cerebrospinal fluid（CSF）in two-dimensional electrophoresis（2-DE）analysis.Methods The CSF sample pool was concentrated to an appropriate volume by ultra-filtration,and then high-abundant proteins were depleted with the commercial Albumin and IgG Removal Kit designed for serum.The remaining CSF containing low-abundant proteins was precipitated by acetone and used for 2-DE analysis and 2-DE Western blot identification.Results Most high-abundant proteins could be depleted with this method.The following 2-DE analysis revealed that low-abundant proteins were obviously enriched after the depletion of high-abundant proteins.Compared with the results of 2-DE Western blot,depleting high-abundant proteins enabled the 3 different fragments of tetranectin to be visible in the 2-DE map.Conclusion The high-abundant protein depletion method for CSF established in this study is simple to perform,and can be helpful in the detection of low-abundant proteins in 2-DE analysis.