| 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 |
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| 引用本文： | 吴本传,陈昭烈,李世崇,刘红,林福玉,张培,黄培堂.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[J].生物技术通讯,2000,11(3):199-201. |
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| 作者姓名： | 吴本传 陈昭烈 李世崇 刘红 林福玉 张培 黄培堂 |
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| 作者单位： | 军事医学科学院生物工程研究所,北京,100071 |
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| 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－Ｓｅｐｈａｒｏｓｅ４Ｂ柱亲和吸附色谱纯化后，最后产品的比性达到６０００００ＩＵ／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了质量ｒｔ－ＰＡ蛋白，ＨＰＬＣ分析达到色谱纯，Ｎ端１５个氨基酸序列和ｐＩ与献报道的一致。蛋白质印迹实验证实具有ｔ－Ｐ
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| 关 键 词： | 组织型纤溶酶原激活剂 纯化 性质 |
Large scale purification of recombiant tissue-type plasminogen activitor and its partial characterization |
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| Wu Benchuan,Chen Zhaolie,Li Shichong,Liu Hong,Lin Fuyu,Zhang Pei,Huang Peiang.Large scale purification of recombiant tissue-type plasminogen activitor and its partial characterization[J].Letters in Biotechnology,2000,11(3):199-201. |
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| Authors: | Wu Benchuan Chen Zhaolie Li Shichong Liu Hong Lin Fuyu Zhang Pei Huang Peiang |
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| Abstract: | 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. |
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| Keywords: | recombinant tissue-type plasminogen activitor purification characterization |
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