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標題: 基因重組蛋白於氫氧基磷灰石固定化金屬離子親和層析法吸附行為之研究
Adsorption behaviors of recombinant proteins on hydroxyapatite- based immobilized metal ions affinity chromatography method
作者: 楊怡馨
Yang, Yi-Sin
關鍵字: hydroxyapatite;氫氧基磷灰石;immobilized metal ions affinity chromatography;IMAC;固定化金屬離子親和層析法
出版社: 化學工程學系所
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本研究利用氫氧基磷灰石(hydroxyapatite)作為固定化金屬離子親和吸附材,以鐵離子Fe3+為配位金屬離子,了解蛋白質在固定化金屬離子親和層析法吸附行為,再利用各個不同恆溫吸附模式分析實驗數據。實驗結果發現其吸附結果較符合Langmuir-Freundlich model,標的蛋白吸附行為則為正合作關係(positive cooperation)。最大吸附量及鍵結親和程度則與分子量及聚組織胺酸標籤數目有關。
在變性狀態(denaturing condition)下,由於蛋白質為未折疊狀態,且所使用的標的蛋白單體分子量相同,所以最大吸附量及解離常數相同。又因分子量較native狀態時分子量小,因此吸附量較native狀態吸附量大。

The adsorption behaviors of three recombinant proteins containing poly(histidine) tags on hydroxyapatite-based immobilized metal ion affinity chromatography (IMAC) are investigated in this study .The experimental data are well fitted with Langmuir-Freundlich isothermal adsorption model, indicating of positive cooperativity for the adsorption of these model proteins.
The adsorption capacity and the binding affinity of the adsorbent for the model proteins are respectively dependent on the size and the number of poly(histidine) tags of proteins. The adsorption isotherms under denaturing conditions are well fitted with the Langmuir model and Langmuir-Freundlich model. The Scatchard analysis further suggest the homogeneous adsorption of the model protein subunits under denaturing conditions. The binding capacities and affinities under denaturing conditions for the three unfolded protein subunits become essentially identical because the molecular size and number of poly(His) tags of the unfolded polypeptide chains of the three protein subunits are the same.
其他識別: U0005-1508200917250500
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