| 用国产微孔玻璃珠初步纯化重组人尿激酶原 |
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| 引用本文： | 郭志霞,叶建新,张正光,高丽华,黄子才,肖成祖.用国产微孔玻璃珠初步纯化重组人尿激酶原[J].生物技术通讯,1998(2). |
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| 作者姓名： | 郭志霞 叶建新 张正光 高丽华 黄子才 肖成祖 |
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| 作者单位： | 军事医学科学院生物工程研究所，军事医学科学院生物工程研究所，军事医学科学院生物工程研究所，军事医学科学院生物工程研究所，军事医学科学院生物工程研究所，军事医学科学院生物工程研究所 北京 100071，北京 100071，北京 100071，北京 100071，北京 100071，北京 100071 |
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| 摘 要： | 用国产微孔玻璃珠从CD-1工程细胞株培养上清中纯化尿激酶原,探索了微孔玻璃珠结合尿激酶原的条件和洗脱方法,比较了硫酸铵线性梯度洗脱或25％乙二醇脱尿激酶原活性蛋白的结果,最后确定洗脱条件为0.25mol/L Tris、1～2mol/L(NH_4)_2SO_4、pH9.3。经此一步纯化,尿激酶原比活性达到15329IU/mg,回收率为78％,还原条件下SDS-PAGE分析分子量为52×10~3的单链尿激酶原占74％。
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| 关 键 词： | 尿激酶原 微孔玻璃珠 硫酸铵 乙二醇 |
Purification of recombinant human prourokinase using domestic micropore glass beads |
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| Guo Zhixia,Ye Jianxin,Zhang Zhengguang,Gao Lihua,Huang Zicai,Xiao Chengzu.Purification of recombinant human prourokinase using domestic micropore glass beads[J].Letters in Biotechnology,1998(2). |
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| Authors: | Guo Zhixia Ye Jianxin Zhang Zhengguang Gao Lihua Huang Zicai Xiao Chengzu |
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| Abstract: | Prourokinase (pro-UK) from the conditioned medium of the genetically-engineered CD-1 cell line was purified using domestic micropore glass beads CMPG) ?and the conditions for the MPG combining pro-UK and the elution process were studied. The elution conditions were confirmed by comparing the effects of the active protein of pro-UK purified by linear gradient elution with ammonium sulfate and by step elution with 25% glycol ethylene, respectively. The results showed that the elulion condition of 0. 25 mol/L Tris, 1-2 mol/L ammonium sulfate with pH.9. 3 was better. The specific activity of purified pro-UK reached 15329 IU/mg, and the recovery rate was 78%. The molecular weight of the final products was 52 kD and the single-chain pro-UK accounted for 74% revealed by SDS-PAGE under reduction conditions. |
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| Keywords: | prourokmase micropore glass beads ammonium sulfate glycol ethylene |
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