研究生: |
邱俊瑋 Jiun-Wei Chiu |
---|---|
論文名稱: |
含聚乙二醇雙性高分子之合成與鑑定以及經酵素固定化後之高分子在水溶液中之自組裝行為 Synthesis and Characterization of Poly(ethylene glycol)-Containing Amphiphilic Graft Copolymer Used as Enzyme Immobilization Carrier and its Self-Assembled Behavior in Aqueous Phase |
指導教授: |
陳崇賢
Chorng-Shyan Chern |
口試委員: |
李振綱
Cheng-Kang Lee 戴子安 Chi-An Dai |
學位類別: |
碩士 Master |
系所名稱: |
工程學院 - 化學工程系 Department of Chemical Engineering |
論文出版年: | 2007 |
畢業學年度: | 95 |
語文別: | 中文 |
論文頁數: | 147 |
中文關鍵詞: | 雙性高分子 、聚乙二醇 、胰凝乳蛋白脢 、酵素固定化 |
外文關鍵詞: | amphiphilic copolymer, mPEG, ChT, enzyme immobilization |
相關次數: | 點閱:296 下載:6 |
分享至: |
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本研究以酯交換反應合成含有聚乙二醇之雙性高分子(PNAS-mPEG),以作為酵素(α-chymotrypsin)固定化載體用。先以傅立葉紅外線光譜(FTIR)進行官能基團鑑定,由核磁共振光譜儀(1H-NMR)計算PEG之接枝效率,進而求出相對平均分子量,並以凝膠滲透層析儀(GPC)與SDS-聚丙烯醯胺膠體電泳(SDS-PAGE)觀察α-ChT是否成功接枝於載體上?
此polymer-protein conjugate系統具有在不同pH值水溶液中的自組裝行為,由螢光光譜、動態光散射光譜儀(DLS)與石英晶體微量天平(QCM)分析產物結構形態與親疏水性。在pH 4的緩衝溶液中,產物有較低的CAC ( 0.8314~2.072 g/l )、較低的2-AN λmax ( 416~422 nm)、較小的粒徑 ( P5-ChT(53.6 nm))與較高的振幅( P10-ChT(148.77 Hz))。在pH 7的緩衝溶液中,則呈現相反之性質。在活性分析上,隨PEG含量之增加其剩餘活性相對提高,且於pH 9下有明顯較高的活性存在。
Amphiphilic copolymers comprising poly(acrylic acid) (PAAc) as the backbone and monomethoxy poly(ethylene glycol) (mPEG) as the grafts were synthesized and characterized. The functional group, grafting efficiency and relative average molecular weight were determined by FTIR, 1H-NMR and GPC, respectively. This copolymer as a carrier is capable of immobilizing chymotrypsin (ChT) in alkaline aqueous phase. GPC and SDS-PAGE were used to whether ChT was success fully attached on to the amphiphilic polymer carrier.
Characteristic properties of polymer-protein conjugate system were analyzed by fluorescence spectroscopy, dynamic light scattering (DLS) and quartz crystal microbalance (QCM). The carrier system has lower CAC values (0.831~2.072 g/l), lower λmax (416~422 nm) of 2-AN spectrum, smaller particle sizes (only for P5-ChT (53.6 nm)) and higher frequency (only for P10-ChT (148.77 Hz)) in pH 4 buffer solutions. On the other hand, this carrier system show opposite results in pH 7.5 buffer solutions. In activity assay, residual activity increased with increasing the mPEG amounts and pH values. This carrier system has obviously high activity in pH 9 buffer solution.
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