中文摘要：

目的：比较不同磁珠及超滤离心管对血清小分子蛋白的分离提纯能力,筛选较优的分离提纯方法。方法：分别利用Cu2＋螯合磁珠（MB-IMAC Cu）、弱阳离子磁珠（MB-WCX）、反相C18磁珠（MB-RPC18）及3种不同型号的超滤离心管对同样4例血清标本进行血清小分子蛋白的分离提纯,比较6种方法对血清高丰度大蛋白的去除能力和处理后血清小分子蛋白质谱图的质量。结果：6种不同的方法均有明显对血清高丰度蛋白的去除能力,其中以超滤离心管效果最明显。在血清小分子蛋白质谱图质量上,磁珠处理后血清生成的质谱图质量更好,其中MB-WCX处理后血清质谱图中总峰数量最多,且在各分子量范围均分布均匀,适于后期鉴定。结论：利用MB-WCX对原始血清进行分离提纯能够获得质量较好的血清小分子蛋白质谱图,可以为进一步实验中寻找差异小分子蛋白并实现兴趣蛋白鉴定奠定良好基础。

英文摘要：

Objective：To screen optimum serum low mass weight（LMW） protein separation and purification methods from six different methods.Methods：Serum samples from four healthy volunteers were extracted by either immobilized metal-affinity chromatography containing copper ions（IMAC-Cu）,weak cation exchange（MB-WCX）,reverse phase C18 magnetic beads（MB-RPC18）or three types of centrifugal filter devices with a MWCO of 3 kDa,10 kDa,30 kDa.Serum LMW protein spectra were generated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF-MS）and analyzed with flexAnalysisTM and Clinprotools 2.1 software and 12% Bis-Tris gel,including the capacity of wiping off high abundance proteins with high mass weight and the quality of serum LMW protein spectra.Results：All of these six ways of protein extraction can wipe off high abundance proteins with high mass weight remarkably and the centrifugal filter devices do the best.However,magnetic beads offer better spectra quality.MB-WCX can provide the most peak amount which distributed homogeneously and got high peak intensity.Conclusions：This study indicates that the serum LMW protein extraction method using MB-WCX can offer serum LMW protein spectra with good quality and will provide sound foundation in further investigation in different serum LMW protein selection and identification.