人血浆载脂蛋白E的分离及纯化

A METHOD FOR ISOLATION AND PURIFICATION OF HUMAN PLASMA APOLIPOPROTEIN E

本文建立了一种快速、简便、较大量分离载脂蛋白Ｅ（ＡｐｏＥ）方法。其优点：①选用高脂血症病人血浆，因ＡｐｏＥ的含量高，来源方便，故克服了分离提纯ＡｐｏＥ时因含量少，不易分离提纯的困难；②采用不连续密度梯度直立头超速离心法，此法只需加两种不同密度的溶液，比经典地超速离心法减少两种不同密度的溶液，故操作方便，且离心时间比经典方法（２４小时）缩短了十分之九；③获得极低密度脂蛋白中的载脂蛋白（ＡｐｏＶＬＤＬ），用ＳｅｐｈａｄｅｘＧ－２００柱层析（１．１×１００ｃｍ）分离提纯得到的ＡｐｏＥ，即是纯的ＡｐｏＥ，且避免了柱太长、不便操作的麻烦。纯化了的ＡｐｏＥ在ＳＤＳ－ＰＡＧＥ中显示一条带，测得分子量为３．８ｋＤａ。

A method was developed for simple and rapid isolation of apolipoprotein E(Apo E)in largquantities. This method is better than other ones. The method has many advantages:①Hyperlipoproteinmichuman plasma was used.It has higher content of Apo E and easier source.②Very low density lipoprotein(VLDL)was prepared by discontinuous density gradient ultracentrifugation in a vertical rotor.The operationof this method was much easier than others. There was only one-tenth as much as others. ③VLDLapolipoprotein(Apo VLDL)was applied to sephadex G-200 chromatography column(1.1×100 cm)forpurification of Apo E. It overcame difficulties of long column.Purified Apo E was identified only one band inSDS-PAGE. The molecular weight of Apo E is 3. 8 kDa.