【摘要】 目的:建立寻常型银屑病血清蛋白组学研究中双向凝胶电泳-质谱技术的实验方法与优化条件。方法:应用ProteoExtract Albumin/IgG Removal Kit纯化血清蛋白,去除白蛋白和IgG,采用固相pH梯度(IPG)、等电聚焦(IEF)为第一向、SDS-PAGE垂直电泳为第二向的双向凝胶电泳(two-dimen-sional gel electrophoresis简称2-DE)和质谱分析评价血清蛋白纯化效果。同时对标本预处理、水化、等电聚焦、SDS-PAGE等步骤进行了优化。结果:采用优化的双向凝胶电泳技术获得了重复性良好的图谱结果,2-DE图谱中分子量在20-55kDa以及pH在4-7区域之间的蛋白点分离效果较好。结论:采用优化后的双向凝胶电泳-质谱技术能够进行寻常型银屑病血清蛋白组学研究。更多还原

【Abstract】 Objective:To establish and optimize the research methods for proteomics analysis of human psoriasis vulgaris by two-dimensional polyacrylamide gel electrophoresis-mass spectrometry.Methods:Removing serum album and IgG was achieved by filtering with ProteoExtract Albumin/IgG Removal Kit,two-dimensional gel electrophoresis(2-DE)technique,including isoelectric focusing(IEF)with an immobilized pH gradient(IPG)and SDS-PAGE was used,associated with mass spectrometry(MS)to evaluate the efficiency of purification.Sample preparation,rehydration,SDS-PAGE,and other steps were adjusted and optimized.Results:The better reproducible results were acquired using the optimized 2-DE technique.The protein spots which locate the area of Mr 20-67 kDa and pH 4-8 were separated well.Conclusions:It is hopeful to carry out serum proteomics research for human psoriasis vulgaris by using optimization of 2-DE-MS technique.更多还原