Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3504
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dc.contributor.advisor孫幸宜zh_TW
dc.contributor.author吳俊逸zh_TW
dc.date2002zh_TW
dc.date.accessioned2014-06-06T05:32:03Z-
dc.date.available2014-06-06T05:32:03Z-
dc.identifier.urihttp://hdl.handle.net/11455/3504-
dc.description.abstractIn this study, commercial regenerated cellulose membranes were modified with different chelating agents to prepare the immobilized metal affinity membranes. The relevant immobilized copper amounts and batch adsorption capacities of different proteins, lysozyme, γ-globulin and BSA, were measured and the effects of pH and salt concentration on adsorption were investigated. The results show that the use of IDA achieved both the largest immobilized amount of copper ions and the largest protein adsorption capacity. As to the pH effect, the protein adsorption was dominated mainly by the affinity binding when the pH values were lower than the pKa for deprotontation of histidine residue on protein molecule surface. On the other hand, at the pH values higher than the pKa, the electrostatic interaction between protein and the nonspecific binding sites on the membranes became significant, but it could be suppressed by adding an optional salt concentration in the solution (0.5 M in this work). For the batch adsorption of three-protein mixture, γ-globulin exhibited the greatest adsorption affinity, whereas lysozyme had the largest adsorption capacity. In the flow experiments, the three proteins could not be separated because all of them had affinities with the immobilized metal affinity membranes.en_US
dc.description.abstract本研究以再生纖維素薄膜為固體載體,改質接上不同之螯合劑,探討其在不同pH值與鹽類濃度下,對於lysozyme、γ-globulin及BSA (bovine serum albumin)的吸附情形。在單一蛋白質的批式吸附部分,以IDA (iminodiacetic acid)為螯合劑所得之銅離子固定量與蛋白質吸附量均是四種螯合劑中最好的。當吸附環境的pH值改變從5.4提高至7.4時,飽和吸附量也隨著提高,顯示吸附受histidine氨基酸殘基上去質子化的親和配位鍵結力影響較大;而當pH值從7.4提高至9.4時,非特定吸附的靜電作用力對吸附效果影響增大。此外,以靜電吸引力為主之非特定性吸附現象可藉加入適當之鹽類濃度(約0.5 M KCl)來改善。在三蛋白質之吸附平衡實驗部分,γ-globulin的吸附力最強,而lysozyme之吸附量最大,均與單一蛋白質之情形相同。但流動吸附實驗之結果則顯示:此三蛋白質均會吸附,故無法有效分離。zh_TW
dc.description.tableofcontents目錄 頁次 中文摘要…………………………………………………………………………….Ⅰ 英文摘要…………………………………………………………………………….Ⅱ 誌謝………………………………………………………………………………….Ⅲ 目錄……………………………………………………………………..…………. Ⅳ 表目錄……………………………………………………………………………….Ⅷ 圖目錄…………………………………………………………………….…………Ⅸ 第一章 前言………………………………………………………….………………1 第二章 文獻回顧……………………………………………………………….……5 2.1固定化金屬親和方法簡介……….……………………………………………5 2.2固體相的組成………………………………………………………………….6 2.3影響吸附的因素………………..………………………………………………8 2.3.1固體載體的種類……………………………………………………….....8 2.3.2螯合劑種類……………………………………………………………….9 2.3.3金屬離子………………………………………………………………….9 2.3.4蛋白質結構…………………………….………………………………..10 2.3.5溫度.……………...……………………………………………………...11 2.3.6 pH值…………………………………………………………………….11 2.3.7鹽類種類及濃度………………………………………………………...12 2.3.8其他因素………………………………………………………………...13 2.4脫附方法………………………..……………………………………………..13 第三章 實驗方法……………………….…………………………………………..15 3.1材料……………………………………………………………………………15 3.2固定化金屬親和薄膜的製備………………….……………………………...15 3.2.1以IDA為螯合劑………………………………………………………..15 3.2.2以TED為螯合劑……………………………………………………….16 3.2.3以Cibacron blue 3GA為螯合劑………………………………………..17 3.2.4以Cibacron red 3BA為螯合劑…………………………………………17 3.3螯合劑及銅離子鍵結量的量測………………………………………………17 3.3.1螯合劑鍵結量的量測…………………………………………………...18 3.3.1.1元素分析儀………………………………………………………18 3.3.1.2紫外/可見光譜儀………………………….……………………..18 3.3.2銅離子鍵結量的量測…….………………………….………………….18 3.3.2.1原子吸收光譜儀…………….…………….……………………..19 3.3.2.2紫外/可見光譜儀………………………………………………...19 3.4蛋白質之吸附實驗.…………………………………………………………...19 3.4.1單一蛋白質………….…………………………………………………..19 3.4.1.1批式吸附平衡實驗………………………………………………19 3.4.1.2非特定性吸附之實驗.……………….…………………………..21 3.4.2三蛋白質…………….…………………………………………………..21 3.4.2.1批式吸附平衡實驗………………………………………………21 3.4.2.2流動吸附實驗……………………………………………………22 3.5液相層析法-混和液中個別蛋白質含量之分析……...……………………...23 第四章 結果與討論…………………….…………………………………………..24 4.1固定化金屬親和薄膜的製備與鍵結量的結果……………………………...24 4.1.1以IDA為螯合劑……………………………….………………………..24 4.1.2以TED為螯合劑……………………………………………………….25 4.1.3以Cibacron blue 3GA為螯合劑………………...……………………...26 4.1.4以Cibacron red 3BA為螯合劑…………………………………………27 4.2單一蛋白質之結果……………………………………………………………28 4.2.1親和吸附等溫線…………………………………………………….......28 4.2.2吸附及脫附時銅離子脫落的影響...……………………………………30 4.2.3非特定性吸附…………………………………………………….……..31 4.2.4 pH值的影響……………………..………………………………….…..33 4.2.5鹽類濃度的影響…………...……………………………………………34 4.3三蛋白質之結果….…………………………………………………………...35 4.3.1批式吸附平衡結果….…………………………………………………..35 4.3.2 流動吸附結果……....………………………………………………….36 第五章 結論…………………………….…………………………………………..37 參考文獻…………………...………………………………………………….…….38zh_TW
dc.language.isozh_TWzh_TW
dc.publisher化學工程學系zh_TW
dc.subject固定化金屬zh_TW
dc.subject薄膜zh_TW
dc.subject再生纖維素zh_TW
dc.title以再生纖維素為材質之固定化金屬親和薄膜的製備與性質之探討zh_TW
dc.typeThesis and Dissertationzh_TW
item.fulltextno fulltext-
item.languageiso639-1zh_TW-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.grantfulltextnone-
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