胆固醇酯转运蛋白的提纯和鉴定

Purification and identification of cholesteryl ester transfer protein

目的：分离纯化人血清胆固醇酯转运蛋白（ＣＥＴＰ），为制备ＣＥＴＰ单克隆抗体和ＣＥＴＰ浓度测定提供物质基础。方法：制备无脂血清，用Ｂｕｔｙｌ－Ｓｅｐｈａｒｏｓｅ４ＦＦ、ＣＭ－ＳｅｐｈａｒｏｓｅＦＦ和ＳｅｐｈａｃｒｙｌＳ－２００三步柱层析分离、纯化ＣＥＴＰ。结果：纯化的ＣＥＴＰ经ＳＤＳ－ＰＡＧＥ与ＣＥＴＰ特异性单克隆抗体免疫反应和活性测定等鉴定证实为纯化ＣＥＴＰ。结论：经三步柱层析获得了纯化ＣＥＴＰ，得率约２５％，活性３０ｎｍｏｌ／（μｇ·ｈ）。

Objectives: To separate and purify human serum choleseryl ester transfer protein (CETP). Methods: Preparation of lipoprotein free serum, purification of CETP by Butyl sepharose 4FF、CM Sepharose FF and Sephacryl S 200 column chromatographic proceduces. Results : This purified protein was confirmed as a human CETP by SDS PAGE、specific immunoreaction with CETP monoclonal antibodies and CETP activity determination. Conclusions: The human serum purified CETP was gained, the activity was 30 nmol/(μg·h). The recovery rate was about 25%. It contributed a base to preparation of CETP monoclonal antibody and determination of CETP concentration.