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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 PA的抗原性
The supernatant of CHO engineering cells producing recombinant tissue-type plasminogen activator (rt PA) was collected and purified by Streamline expanded bed and lysine Sepharose 4B column affinity chromatography. The specific activity of the final product reached 6 The recovery of total activity of rt PA was about 98% at 0 0 0 0 0 mg / L. The main relative high molecular weight rt PA was analyzed by SDS-PAGE. The chromatographic purity and N-terminal 15-amino acid sequence were obtained by HPLC. Reported the same. Western blotting confirmed t PA antigenicity