聚焦电层析分离牛血清白蛋白和牛血红蛋白

Separation of Bovine Serum Albumin and Bovine Hemoglobin by Focusing Electrochromatography

本文借鉴聚焦层析的原理,研究聚焦电层析分离牛血清白蛋白和牛血红蛋白的方法。在充填阴离子交换葡聚糖凝胶DEAE-Sephadex A-50的柱中加入pH5.7缓冲液平衡,再加入pH3.0缓冲液形成pH梯度。在柱的两端施加电场,正极在上端,负极在下端。蛋白质在pH梯度内移动时,所带电荷发生变化,所受电场力也改变,移动速度的差异增大,牛血清白蛋白和牛血红蛋白可以完全分离,回收率为90%以上。操作时要注意调节pH梯度范围,以避免蛋白质沉淀析出。这种方法简便,价廉,适用于分离等电点不同的蛋白质。

A focusing electrochromatography method was developed to separate the mixture of bovine serum albumin and bovine hemoglobin according to the principle of chromatofocusing. Anion exchange gel DEAE Sephadex A-50 was packed in a column. A buffer （pH 5.7） was added into the column for equilibrium, then another buffer （pH 3.0） was added to form pH gradient. Electric field was applied on two ends of the column. The upper electrode was anode and the lower one was cathode. During moving course of proteins in pH gradient, the electric charge of proteins changed, and the electric field force on them also altered, so the difference of moving velocity between bovine serum albumin and bovine hemoglobin was enhanced. As a result, the proteins were separated thoroughly and the recovery yields were over 90%. The range of pH gradient should be adjusted to aviod the precipitation of protein. This method was simple and cheap, which was suitable to separate proteins with different isoelectric points.