重组组织型纤溶酶原激活剂大规模纯化及部分性质的研究  

Large scale purification of recombiant tissue-type plasminogen activitor and its partial characterization

作　　者：吴本传[1] 陈昭烈[1] 李世崇[1] 刘红[1] 林福玉[1] 张培 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2000年第3期199-201,共3页Letters in Biotechnology

摘　　要：收集产重组组织型纤溶酶原激活剂 (rt PA)的CHO工程细胞灌流培养上清 ,经过Streamline扩张柱床和赖氨酸 Sepharose 4B柱亲和吸附色谱纯化后 ,最后产品的比活性达到 6 0 0 0 0 0IU mg蛋白 ,rt PA总活性回收率在 98%左右 ,经还原SDS PAGE分析主要为高相对分子质量rt PA蛋白 ,HPLC分析达到色谱纯 ,N端 15个氨基酸序列和pI与文献报道的一致。蛋白质印迹实验证实具有tTissue-type plasminogen activitor produced by recombiant CHO cell line was purified from large volumes of supernatant fluid using a reusable continuous chromatography system comprising streamline and Lysine-Sepharose 4B.The final product had a specific activity of about 600 000 IU/mg protein,total activity yield was about 98%,HPLC showed chromatogrphic purity.Analysis by SDS-PAGE in the presence of reducing agents and by staining with Coomassie brilliant blue R250 showed one main band with a high molecular weight.The isoetectric point and N-terminal amino acid sequence of the rt-PA correspond to others references,Western blotting analysis showed that the product has natural t-PA antigen characterization.

关 键 词：组织型纤溶酶原激活剂 纯化 性质 

分 类 号：TQ464[化学工程—制药化工] R973.2[医药卫生—药品]