博碩士論文 84341002 詳細資訊




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姓名 蔡宜勳( Yi-Shiun Tsai)  查詢紙本館藏   畢業系所 化學工程與材料工程學系
論文名稱 蛋白質與疏水性吸附劑間交互作用之研究: 鹽濃度與疏水性觸手長度之影響
(Studies of the Interaction between Protein and Hydrophobic Absorbents : Effects of Salt Concentration and Hydrophobic Ligand Chain Length)
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摘要(中) 經由平衡鍵結分析(the equilibrium binding analysis)和ITC直接測定吸附火含,探討硫酸銨(ammonium sulfate,(NH ) SO )鹽濃度在肌紅蛋白(myoglobin)對兩種疏水性吸附劑(hydrophobic adsorbents):butyl-和octyl-sepharose上之吸附反應的影響。由結果顯示肌紅蛋白與兩種疏水性吸附劑的吸附親和力,會隨著鹽濃度的增加而變強;再者,由恆溫滴定微卡計(isothermal titration calorimetry,ITC)所測定的結果,顯示,吸附火含的變化量隨鹽濃度的增加而減少,這個發現可被解釋為:隨鹽著濃度的增加去水合熱(dehydration heat)會減少且由於疏水性反應的關係,釋放熱因此增加。此外,隨著鍵結蛋白質的增加,兩種樹脂對肌紅蛋白的吸附火含皆會隨著增大,而且其變化程度在不含(NH ) SO 溶液中比在1.0M (NH ) SO 中顯著,說明了,蛋白質間電性斥力在無鹽的狀態下對於蛋白質吸附量影響是較強的。
在此,所討論的熱力學參數是有其重要含意的,尤其是吸附焓與吸附火商的相對變化量,不只對蛋白質吸附現象的機制提供更細微的內視,而且對HIC也有更進一步的理論分析。
摘要(英) The effect of (NH4)2SO4 concentrations on the interaction mechanism between myoglobin and two hydrophobic adsorbents, butyl- and octyl-Sepharose was investigated by the equilibrium binding analysis and by directly measured adsorption enthalpies. The result obtained from the isotherms demonstrated that the affinities of myoglobin adsorption onto the both adsorbents were increased with salt concentrations. Furthermore, the adsorption enthalpies measured by using isothermal titration calorimetry (ITC) were decreased with an increment of salt concentrations, and these findings were explained by the reduction in the dehydration heat and the enhancement of heat released by the hydrophobic interaction as salt concentrations increase.
Additionally, the adsorption enthalpies of myoglobin with both the resin increased as the amount of bound protein, and the increment in the variation of enthalpy value at solution without (NH4)2SO4 appeared to be steeper than that at 1.0 M (NH4)2SO4. This result implies that the effect of protein-protein electric repulsion on the amount of bound protein is stronger in the absence of the salt in this experiments.
The thermodynamic parameters presented herein have important implication, particularly the relative variation between the adsorption enthalpies and the adsorption entropies, both for providing further insight into the binding mechanism of protein adsorption and for improving theoretical approaches to HIC.
關鍵字(中) ★ 恆溫滴定微卡計
★ 蛋白質吸附
★ 層色分析
★ 疏水性交互作用
★ 焓變化
★ 蛋白質純化
關鍵字(英) ★ Hydrophobic interaction chromatography
論文目次 封面
中文摘要
英文摘要
目錄
圖目錄
表目錄
第一章 緒論
第二章 文獻回顧
2.1 液相層析法之發展
2.2 離子交換層析(Ion Exchange Chromatography, IEC)
2.3 疏水交互作用層析(Hydrophobic Interaction Chromatography, HIC)
2.4 親和力層析法(Affinity Chromatography, AC)
2.5 以分子大小分離層析法( Size Exclusion Chromatography,SEC )
2.6 鹽濃度與觸手長度的影響
2.7 熱力學探討
第三章 研究目的
第四章 理論基礎
4.1 疏水性交互作用層析法的吸附機制
4.2 影響吸附機制的因素
4.3 HIC的熱力學探討
4.5 恆溫滴定微卡計
4.6 卡計與Van't Hoff方程式所得焓值之差別
4.7 本研究立論原理
第五章 儀器設備與藥品
5.1 實驗藥品
5.2 儀器設備
第六章 實驗步驟與分析
6.1 平衡鍵結等溫吸附線的量測
6.2 吸附焓變化的量測
第七章 結果與討論
7.1 平衡鍵結分析
7.2 微卡計測量
第八章 結論
第九章 參考文獻
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指導教授 周正堂(Cheng-tung Chou) 審核日期 2001-7-17
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