寻常型银屑病血清蛋白质组学研究中双向凝胶电泳-质谱技术的初步建立  

Preliminary establishment of two-dimensional gel electrophoresis-mass spectrometry for serum proteomics analysis in psoriasis vulgaris

作　　者：刘占奎[1] 谭升顺[1] 于春水[1] 樊靖华[1] 白转丽[1] 李俊杰[1] 

机构地区：[1]西安交通大学第二附属医院皮肤科,西安710004

出　　处：《中国麻风皮肤病杂志》2008年第4期264-266,共3页China Journal of Leprosy and Skin Diseases

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。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 ProteoExtraet Albumin/IgG Removal Kit, two- dimensional gel electrophoresis（2- DE） technique, including isoclectric 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.

关 键 词：蛋白组学 双向凝胶电泳 质谱 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]