两步柱色谱法分离纯化重组肿瘤血管生长抑制因子Kringle 5  

Two-Step Chromatographic Method for the Separation and Purification of Recombinant Angiogenesis Inhibitor Kringle 5

作　　者：贾信贵[1] 边六交[1] 

机构地区：[1]西北大学生命科学学院国家微检测系统工程技术研究中心,陕西西安710069

出　　处：《色谱》2007年第3期344-347,共4页Chinese Journal of Chromatography

基　　金：陕西省科技攻关项目(No.2003K10-G58)

摘　　要：建立了一种用Ni2+螯合的Chelating Sepharose Fast Flow亲和柱色谱和Sephadex G-75凝胶排阻柱色谱分离纯化重组肿瘤血管生长抑制因子Kringle5的方法。采用该工艺得到的重组Kringle5经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明其纯度约为98%,且具有抑制鸡胚绒毛尿囊膜新生血管生成的生物活性。The Kringle 5 domain of plasminogen, which was previously shown to inhibit angiogenesis in vitro and in vivo, is one of the most potent angiogenesis inhibitors known to date. A two-step chromatographic method, which consists of Chelating Sepharose Fast Flow chelated with Ni^2＋ affmity medium and Sephadex G-75 medium, was established to separate and purify recombinant angiogenesis inhibitor Kringle 5 （rK5）. Through the two-step chromatographic purification process, the obtained rK5 was confirmed to be homogeneous on sodium dodecyl sulfatepolyacrylamide gel electrophoresis （SDS-PAGE） and its relative molecular mass was estimated to be about 32 000, which matched well with the prediction by gene sequence. Its purity was about 98%, and the total protein recovery of this method was 0.63%. In addition, it was found that it inhibited the blood vessel growth of chick embryo chorioallantoic membrane effectively.

关 键 词：亲和柱色谱法 凝胶排阻柱色谱法 重组血管生长抑制因子Kringle 5 分离 纯化 

分 类 号：O658[理学—分析化学]