大豆脂肪氧化酶同功酶抗原的快速纯化方法  

Rapid Approach of Soybean Lipoxygenase'antigen Purification

作　　者：马志民[1] 刘渊[1] 蒋春志[1] 杨春燕[1] 马峙英[1] 张孟臣[1] 

机构地区：[1]国家大豆改良中心石家庄分中心

出　　处：《大豆科学》2008年第2期306-309,共4页Soybean Science

基　　金：河北省自然科学基金资助项目(C2005000605)

摘　　要：目前常用的蛋白纯化方法均存在操作复杂、纯化周期长、成本高等缺点,本试验试图探索出一种经济快捷的蛋白纯化方案,且纯化后的蛋白可以用于制备单克隆抗体;以普通大豆品种冀豆12以及脂肪氧化酶缺失近等基因系Suzuyutaka中的Su-1,3,Su-2,3为材料,将大豆脂氧酶非变性PAGE分析与电洗脱技术相结合,纯化脂肪氧化酶同功酶抗原蛋白;回收到具有活性的电泳纯级脂肪氧化酶溶液,回收率达到了与离子交换柱层析相当的水平(49.64%);本试验探索的方法实现了应用经济快捷的纯化方案获得免疫所需抗原,为尽早获得脂肪氧化酶单克隆抗体并将其应用于实践打下了基础,并为其它蛋白质的快速分离纯化提供了参考。The common method of purifying protein has the shortcoming of complex operation,time- consuming of purifying, high costing and so on. The objective of this paper is to explore a fast and economical process of protein purification,and the purified protein can be applied to prepare anti-this protein monoclonal antibody. Combining both Native PAGE of the soybean Lipoxygenase and Electro Elution,we purified the lipoxygense from the soybean Jidou No. 12 and the Su-1,3, Su-2,2. Lipoxygenase solution is retrieved with electrophoresis degree and recovery rate 49.64% as same of the ion exchange column chromatography. This built a fast and economical approach for antigen purifying,and it is helpful for preparing anti-lipoxygenase monoclonal antibodies and applied to practice even. It also offers important reference to the purification of other protein.

关 键 词：脂肪氧化酶 非变性PAGE 电洗脱 纯化 

分 类 号：S565.1[农业科学—作物学] TS202.3[轻工技术与工程—食品科学]