LpB:E标准物的制备

THE PREPARATION OF STANDARD FOR LpB:E DETERMINATION

用纯化的人ＬＤＬ（ａｐｏＢ）免疫山羊，获得特异性羊抗人ａｐｏＢ抗血清。将用正辛酸、硫酸铵沉淀和Ｐｒｏｔｅｉｎ－Ｇ亲和层析纯化的羊抗人ａｐｏＢＩｇＧ与ＣＮＢｒ－ａｃｔｉｖａｔｅｄＳｅｐｈａｒｏｓｅ４Ｂ共价交联，可制备获得特异性抗体亲和层析柱。将适量的人血清通过该亲和柱，用高ｐＨ缓冲液冲洗后，用低ｐＨ缓冲液洗脱，可获得含ａｐｏＢ的脂蛋白，其中包括了同时含ａｐｏＢ和ａｐｏＥ的ＬｐＢ：Ｅ组分。测定其ａｐｏＥ含量后，该组分可作为测定ＬｐＢ：Ｅ的标准物。

The goat antiserum against human apoB was obtained by immunizing goat with purified human LDL.The goat IgG purified by selective precipitation with caprylic acid and ammonium sulphate and Protein G affinity chromatography was covalently coupled with CNBr activated Sepharose 4B.The lipoprotein particals (LpB:E)containing apolipoprotein B with apolipoprotein E were isolated by an affinity charomatography on antibodies against human apolipoprotein B and used as the standard for LpB:E determination.