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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >Displacement chromatography of proteins on hydrophobic charge induction adsorbent column
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Displacement chromatography of proteins on hydrophobic charge induction adsorbent column

机译:疏水电荷诱导吸附柱上蛋白质的置换色谱

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Displacement chromatography of protein mixtures is proposed on hydrophobic charge induction chromatography (HCIC). We have used an HCIC medium, MEP-Hypercel as the stationary phase and a quaternary ammonium salt, benzethonium chloride, as the displacer. It was found that the multiple interactions between proteins/displacer and the HCIC sorbent, i.e. hydrophobic interaction and charge repulsion, enabled a greater flexibility for the design of displacement processes and ease of column regeneration by adjustment of pH. The capacity factors of proteins and displacers were used to predict their performances in column displacement, and the experimental results agreed well with the prediction. An isotachic displacement train of lysozyme and a-chymotrypsinogen A was formed with benzethonium chloride as the displacer at pH 5.0 with good yields and purities of the two proteins. Column regeneration was efficiently achieved by charge repulsion between the displacer and the adsorbent at lower pH values (pH 3 and 4). The results indicate that the displacement chromatography on HCIC is a good alternative to traditional hydrophobic displacement chromatography. (c) 2007 Elsevier B.V. All rights reserved.
机译:蛋白质混合物的置换色谱法是在疏水电荷感应色谱法(HCIC)上提出的。我们使用了HCIC介质MEP-Hypercel作为固定相,并使用了季铵盐苄索氯铵作为置换剂。已经发现,蛋白质/置换剂与HCIC吸附剂之间的多种相互作用,即疏水性相互作用和电荷排斥,为置换过程的设计提供了更大的灵活性,并通过调节pH使色谱柱易于再生。蛋白质和置换剂的容量因子用于预测其在色谱柱置换中的性能,实验结果与预测结果吻合良好。在pH 5.0时,用苄索氯铵作为置换剂,形成了溶菌酶和α-胰凝乳蛋白酶原A的等速置换序列,两种蛋白质的收率和纯度都很高。通过在较低的pH值(pH 3和4)下置换器和吸附剂之间的电荷排斥有效地实现了色谱柱再生。结果表明,HCIC上的置换色谱法是传统疏水置换色谱法的良好替代方案。 (c)2007 Elsevier B.V.保留所有权利。

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