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大豆科学  2008 

大豆脂肪氧化酶同功酶抗原的快速纯化方法

DOI: 10.11861/j.issn.1000-9841.2008.02.0306, PP. 306-309

Keywords: 脂肪氧化酶,非变性PAGE,电洗脱,纯化

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Abstract:

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。

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