中压液相层析法纯化人血清载脂蛋白AⅠ和AⅡ的研究

Purification of Human Serum Apolipoprotein AⅠand AⅡ by Middle-Pressure Liquid Chromatography

目的 建立中压液相层析分离法纯化人 HDL 主要载脂蛋白 A 、A 等的方法。方法 以人血清高密度脂蛋白制品为原料 ,经一次性密度梯度超离心法纯化 HDL。去脂 HDL 经中压 Sephacryl S- 2 0 0柱层析获apo E、A 、A 及 Cs粗品。用 Sephadex G- 75柱层析分离 apo A 及 A 。结果 纯化的载脂蛋白 apo A 及 A 经免疫和 SDS- PAGE鉴定为免疫纯及电泳纯。结论 建立了一种可大量、快速、高分辨分离纯化载脂蛋白 apo A 及 A 的方法 ,为今后载脂蛋白和脂蛋白的基础和临床研究提供必要的实验材料。

Objective This study is made for the purification of human HDL apolipoproteins by middle-pressure liquid chromatography(MPLC). Methods Human HDL was isolated by one step density ultracentrifugation. Delipided human HDL was separated by Sephacryl S-200 molecular sieve chromatography. Sephadex G-75 molecular sieve chromatography was used to separate apoAⅠ and apoAⅡ. Results Purified apoAⅠ and apoAⅡ were obtained and SDS-PAGE and immunodiffusion test indicated that the proteins are the same as those theoretically predicted. Conclusion We have established a purification procedure for human HDL apolipoproteins with high efficiency and simplicity by MPLC. It could serve as a base for preclinical and clinical trials of HDL apolipoproteins.