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t
Tissue-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.
出处
《生物技术通讯》
CAS
2000年第3期199-201,共3页
Letters in Biotechnology
关键词
组织型纤溶酶原激活剂
纯化
性质
recombinant tissue-type plasminogen activitor
purification
characterization
