胆固醇酯转运蛋白的提纯和鉴定

Purification and identification of cholesteryl ester transfer protein

目的建立一种新的分离纯化人血清胆固醇酯转运蛋白（ＣＥＴＰ）的方法，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ＣＥＴＰ经ＳＤＳＰＡＧＥ电泳鉴定呈单一区带，分子量６５０００，蛋白质含量０３２５ｍｇ／ｍｌ，与国外和作者研制ＣＥＴＰ单抗的免疫斑点试验呈特异性反应，与纯化脂蛋白（ａ）、载脂蛋白（Ａｐｏ）Ａ１、ＡｐｏＢ、ＩｇＧ、ＩｇＡ、ＩｇＥ、Ｃ３、Ｃ４抗体无免疫反应。结论经三步柱层析获得了纯化ＣＥＴＰ，得率约２５％，活性每小时３０ｎｍｏｌ／μｇ。

Objective To separate and purify human serum choleseryl ester transfer protein (CETP). Method Lipoprotein free serum, purification of CETP was completed 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. The activity was 30 nmol/μg/h. The recovery rate was about 25%. Conclusion The method contributes to preparation of CETP monoclonal antibody and determination of CETP concentration.