作　　者： ; ;

机构地区： 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

出　　处： 《色谱》 2007年第3期344-347,共4页

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 分离 纯化

领　　域： [理学] [理学]