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超氧化物歧化酶分离-修饰耦合过程的研究

Study on the coupling procedure of SOD’s purification and modification

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【作者】 陈佳乐高飞易喻应国清

【Author】 CHEN Jia-le1,GAO Fei2,YI Yu1,YING Guo-qing1(1.College of Pharmaceutical Science,Zhejiang University of Technology,Hangzhou 310032,China;2.ABON Biopharm(Hangzhou) Co.,Ltd.,Hangzhou 310018,China)

【机构】 浙江工业大学药学院艾博生物医药(杭州)有限公司

【摘要】 以超氧化物歧化酶(SOD)为模板蛋白,研究其分离—修饰耦合工艺.即将SOD选择性地吸附在铜离子螯合葡聚糖凝胶层析柱上,先不经洗脱分离,直接用活化的聚乙二醇(PEG)5000对吸附的SOD进行化学修饰,再通过特异性洗脱分离获得PEG-SOD,使得SOD分离纯化和修饰过程同步完成.获得了一条SOD金属螯合层析分离—PEG化学修饰耦合工艺路线.获得的PEG-SOD比活9 055.1 U/mg,纯化倍数1.75倍,氨基修饰率42.3%,修饰前后活性保留87.1%.证明SOD的分离-修饰耦合工艺是可行的.

【Abstract】 This article uses SOD as a template,to study its coupling procedure of purification and modification.SOD was mixed with Cu2+ chelating-sephadex G-75 without being washed immediately but modified by activated PEG5000 upon the solid-phase column,then washed specifically.The modification and purification of SOD completed synchronously.The specific activity of the obtained PEG-SOD is 9 055.1 U/mg,with a purification factor of 1.75,modified at 42.3% of its free amino groups,retained 87.1% of enzymatic activity of native SOD.The results confirm that it is feasible to couple the purification and modification of SOD.

【基金】 浙江省重大科技攻关资助项目(2005C13025)
  • 【文献出处】 浙江工业大学学报 ,Journal of Zhejiang University of Technology , 编辑部邮箱 ,2009年01期
  • 【分类号】Q55
  • 【下载频次】223
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