胆固醇酯转运蛋白的提纯和鉴定  

Purification and identification of cholesteryl ester transfer protein

作　　者：庄一义[1] 汪俊军[1] 李勇[1] 

机构地区：[1]南京军区南京总医院全军医学检验中心

出　　处：《中华医学检验杂志》1999年第2期101-103,共3页

基　　金：解放军九五科研基金

摘　　要：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.

关 键 词：胆固醇酯类 载体蛋白质类 提纯 鉴定 CETP 

分 类 号：Q548.103[生物学—生物化学] Q513.503