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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 a method for the removal of high abundance protein in cerebrospinal fluid and to evaluate by two-dimensional electrophoresis. Methods The samples of cerebrospinal fluid were concentrated to the appropriate volume by ultrafiltration and treated with commercial serum albumin and immunoglobulin G (IgG) removal kits to obtain cerebrospinal fluid containing low-abundance proteins, desalted by acetone precipitation, Dimensional gel electrophoresis (2-DE) analysis and 2-DE Western blot identification. Results This method can effectively remove albumin and IgG from cerebrospinal fluid. The results of 2-DE showed that the low abundance protein in cerebrospinal fluid was significantly enriched after removal of high abundance protein. Compared with the results of 2-DE Western blot, A different fragment to remove the high abundance of protein can be fully revealed. Conclusion The high abundance protein removal method of cerebrospinal fluid established in this study is simple and convenient, which is beneficial to the detection of low abundance proteins in two-dimensional electrophoresis.