人血浆载脂蛋白B100分离纯化方法的研究

Study on purification of human ApoB100

通过硫酸右旋糖苷(dextransulfate,DS)法沉淀人血浆中低密度脂蛋白(LDL)及利用SDS-PAGE电泳分离出其中载脂蛋白B100(apolipoprotein B100,ApoB100),与弗氏佐剂充分乳化后免疫新西兰大白兔,获得ApoB100特异性多克隆抗体,经过protein A纯化后制备抗体亲和层析柱,并利用该亲和层析柱分离纯化SDS-PAGE电泳ApoB100条带的电洗脱液和人血浆LDL中ApoB100.结果发现,所制备层析柱对电洗脱液中ApoB100具有较好纯化效果.

Low density lipoprotein （LDL） was run on SDS-PAGE after the preparation of LDL by the dextransulfate （DS） method, and ApoB100 was recovered by using eleetro-eluter. New Zealand white rabbits were immunized with ApoB100. The anti-serum showed special sensitivity to ApoB100 and anti-ApoB100 IgG was isolated from rabbit anti-serum by protein A agarose beads. Using CNBR activated sepharose FF combined with anti-ApoB100 IgG as the immunoaffinity chromatographic column, the separation of ApoB100 from SDS- PAGE or LDL is evidently different.