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疏水层析是分离生物大分子的常用技术之一,但对疏水层析中蛋白质吸附动力学和平衡过程的研究并不多见。本文对蛋白质疏水吸附动力学和平衡过程作了基本假设,并用实验进行了验证。制备了两种不同丁基密度的疏水琼脂糖介质,用其吸附牛血清白蛋白(BSA)以验证对疏水吸附动力学与平衡过程作的假设,考察了盐浓度及配基密度对蛋白质疏水吸附的影响。还对三种疏水性不同的蛋白质:核糖核酸酶、卵清蛋白和牛血清白蛋白的混合体系进行了分离性能的研究,获得了满意的分离效果。实验表明,蛋白质在疏水介质上的吸附动力学和平衡过程与所作假设相符,在实验条件下等温吸附线符合Langmuir吸附等温方程;研制的丁基琼脂糖疏水介质具有优良的使用性能。
Abstract：Hydrophobic Interaction Chromatography(HIC) is widely used in separation and purification of biological macromolecules. The study of adsorption kinetics and equilibrium process of protein in HIC is seldom reported. In this article, basic hypothesis of adsorption kinetics and equilibrium process of protein in HIC was established and verified with self-prepared butyl-agarose uncharged HIC media and bovine serum albumin (BSA). In addition, effects of salt concentration and butyl group density on adsorption of protein in HIC were investigated. Then the media were used to separate three proteins with different hydrophobicity: RNase, ovalbumin and BSA, the satisfied result was obtained. It is shown that adsorption kinetics and equilibrium process of protein in HIC were consistent with the basic hypothesis, the adsorption isotherm well fitted to Langmuir adsorption equation. Self-prepared butyl-agarose HIC media showed good properties for adsorption and separation of protein.
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基本信息:
中图分类号:O629;O658
引用信息:
[1]孟庆强,乔昕,马光辉,郑春,苏志国.疏水层析蛋白质动力学与平衡过程的考察[J].离子交换与吸附,2004(05):391-399.
基金信息:
国家863计划 (2002AA629040);; 国家自然科学基金 (20136020)