中医证侯血浆蛋白质双向凝胶电泳方法的建立及优化

目的建立中医证侯血浆蛋白质双向凝胶电泳（2-DE）方法并对其进行优化。方法以正常人血浆样品为研究对象,对白蛋白/IgG球蛋白去除后血浆样品、全血浆和热SDS法全血浆样品进行pH梯度2-DE分离,凝胶经硝酸银染色后,利用PDQuest软件对2-DE图谱进行重复性分析,并对不同处理方法的全血浆2-DE图像的结果进行比较。结果与全血浆直接上样2-DE凝胶图谱比较,白蛋白/IgG球蛋白去除后血浆上样双向电泳蛋白有所增多,但由于白蛋白/IgG球蛋白去除过程中的非特异性吸附,造成了同一样品在蛋白质图谱上的明显差异;与全血浆直接上样和白蛋白/IgG球蛋白去除后血浆样品上样2-DE凝胶图谱比较,热SDS法处理血浆样品可以得到更多的蛋白质,且重复性好（P〈0.05）,凝胶硝酸银染色显示蛋白质点染色深、分散清楚。结论热SDS法全血浆上样2-DE分析法具有蛋白丢失少、操作简便、重复性好的特性,是证侯血浆蛋白质组学研究的有效手段。

Establish and optimize of plasma protein 2-DE of syndrome

Objective Syndrome of plasma proteins to establish two-dimensional gel electrophoresis and its optimization. Methods Normal human plasmas was used in this study. The samples were treated with three different ways： removing albumin and IgG globulin, boiling and no treating. The samples were then analyzed by two-dimensional gel electrophoresis with different pH gradients. After staining with argenti nitras, the gels were scanned and analyzed by PDQuest software. Results The electrophoregram of normal human plasmas was obtained by two-dimensional gel-electrophoresis. By silver nitrate staining,the proteins SDS hot plasma sample of the entire two-dimensional gel-electrophoresis staining deep, scattered clear. Conclusion Because of little-losing, simple and good repeatability, the method of two-dimensional gel- electrophoresis using heating-SDS whole plasma sample is an effective means of the syndrome plasma proteomics research.