Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/66432
DC FieldValueLanguage
dc.contributor.advisor陳震漢zh_TW
dc.contributor.advisorShinn-Jyh Dingen_US
dc.contributor.advisor丁信智zh_TW
dc.contributor.author李智雄zh_TW
dc.contributor.authorLee, Chih-Hsiungen_US
dc.date2005zh_TW
dc.date.accessioned2014-06-09T09:31:47Z-
dc.date.available2014-06-09T09:31:47Z-
dc.identifier.urihttp://hdl.handle.net/11455/66432-
dc.description.abstract明膠(gelatin)是膠原蛋白經過酸性或鹼性處理後萃取所得的產物,因處理方式差異使明膠有兩種不同的等電點。作為生醫材料的明膠除具有生物降解性、生物相容性、無毒性等優點外,更有比膠原蛋白更低的抗原性。藉由明膠不同等電點的特性可將蛋白質經靜電引力作用而與明膠基質接合在一起,一旦明膠基質在體內經由酵素分解,蛋白質就從明膠基質上釋放出來。 Marty最早提出製備明膠奈米顆粒的方法,之後由Coester等人改良。本研究應用此方法,探討戊二醛和pH值對明膠奈米顆粒性質的影響。先將購得的明膠分子溶於水中並且加入等體積的丙酮使大部分的明膠沉澱,將沉澱的明膠回溶於水中並且調整成不同的pH值,之後加入不同濃度的戊二醛交聯明膠分子以形成不同的明膠奈米顆粒。分析明膠奈米顆粒的形態、結晶性、粒徑、生物降解性、吸附性等特性。X光繞射儀測得明膠膜呈現非晶質結構;掃描式電子顯微鏡顯示明膠奈米顆粒是管狀的,約50×300 nm大小,與實驗條件無關;穿透式電子顯微鏡顯示明膠奈米顆粒是由直徑約30-40 nm小分子凝聚而成的分子團;光散射粒徑分析儀顯示明膠奈米顆粒的範圍在180-374 nm之間,大小受戊二醛濃度和pH值所影響;陽離子交換液相色層分析法顯示明膠奈米顆粒被胰蛋白脢分解;CBD-RGD融合蛋白質與明膠奈米顆粒作用後,SDS-PAGE顯示蛋白質吸附到明膠奈米顆粒上。zh_TW
dc.description.abstractGelatin is a denatured protein obtained by acid or alkaline processing of collagen. These different processing resulted in gelatin with different isoelectric points (IEPs). Gelatin has a number of advantages as a biomaterial. It is biocompatibility, biodegradability and nontoxicity; more importantly, much lower antigenicity than collagen. We can associate the charged proteins to oppositely charged gelatin matrix through isoelectrical nature of gelatin. Thus the protein is accordingly released as gelatin matrix enzymatically degrades in living body. One established method is the precipitation of gelatin particles from gelatin solution using two-step desolvation by Coester et al. following the pioneering studies of Marty and coworkers. The objective of this study was to prepare gelatin particles from gelatin solution using the modified method of two-step desolvation, focusing on the experimental conditions with different gultaradehyde concentration and pH values. First, the as-received gelatin was dissolved in water under gentle heating and the desolvation step was initiated by adding acetone. Sequentially, the solution pH was adjusted to different values and then adding different concentrations of glutaraldehyde. Morphology, crystallinity, particle size, biodegradation, and CBD-RGD adsorption of gelatin nanoparticles formed were evaluated. The experimental results showed that all gelatin membranes were an amorphous phase. The morphologies of all gelatin particles were column-like crystallites with 50300 nm in size under SEM independent on experimental conditions. Transmission electron microscope further indicated the appearances of the gelatin clusters consisting of about 30-40 nm particles. Dynamic light scattering showed the mean cluster (same as particle herein) size is in the range of 180 to 374 nm, consistent with the observation of morphology, relating significantly to glutaraldehyde concentration and pH effect. Cation exchange liquid chromatography showed gelatin nanoparticles were degradable in trypsin. After mixing CBD-RGD fusion proteins with gelatin nanoparticles, SDS-PAGE showed CBD-RGD fusion proteins were absorbed onto gelatin nanoparticles.en_US
dc.description.tableofcontents中文摘要 1 英文摘要 2 第一章 前言及研究目的 3 1-1 前言 3 1-2 研究目的 4 第二章 文獻探討 5 2-1 明膠的來源 5 2-2 明膠的分子量 5 2-3 明膠的等電位點 6 2-4 明膠的免疫性質 6 2-5 明膠的生物適應性 7 2-6 明膠奈米顆粒 8 2-6-1 製備方法 8 2-6-2 奈米顆粒的應用 9 2-7 RGD序列 10 第三章 材料與方法 12 3-1 材料 12 3-2 SDS-PAGE分析明膠原料純度 13 3-3 製備明膠奈米顆粒 14 3-4 CBD-RGD融合蛋白質合成及純化 14 3-4-1 合成RGD蛋白質 15 3-4-2 測定蛋白質濃度 15 3-4-3 陰離子交換液相層析法純化融合蛋白質 16 3-4-4 SDS-PAGE分析純化後的蛋白質溶液 17 3-5 明膠膜評估 18 3-5-1 微結構觀察 18 3-5-2 機械性質 18 3-5-3 相組成 18 3-6 分析明膠奈米顆粒 19 3-6-1 粒徑分析 19 3-6-2 微結構觀察 19 3-6-3 陽離子交換液相層析法純化明膠奈米顆粒 20 3-6-4 胰蛋白脢分解明膠奈米顆粒 21 3-6-5 SDS-PAGE評估RGD與明膠顆粒吸附作用 21 3-6-6 陰離子交換液相層析法評估RGD與明膠顆粒吸附作用22 第四章 結果 23 4-1 SDS-PAGE分析明膠原料純度 23 4-2 明膠奈米顆粒水溶液 23 4-3 明膠奈米顆粒的產率 23 4-4 CBD-RGD融合蛋白質合成及純化 24 4-5 明膠膜的微結構、機械性質及相組成 24 4-6 明膠奈米顆粒的特性 24 4-6-1 粒徑大小 25 4-6-2 微結構 25 4-6-3 陽離子交換液相層析法定性奈米顆粒 26 4-6-4 降解性 26 4-6-5 與RGD融合蛋白質間之吸附作用 26 4-6-5-1 陰離子交換液相層析法 26 4-6-5-2 SDS-PAGE分析過濾液 27 第五章 討論 28 5-1 明膠奈米顆粒的製備 28 5-1-1 SDS-PAGE分析明膠原料 28 5-1-2 明膠溶液 28 5-1-3 產率 30 5-2 CBD-RGD融合蛋白質合成及純化 30 5-3 明膠膜的結構、機械性質及相組成 31 5-4 明膠奈米顆粒的特性 32 5-4-1 粒徑大小 32 5-4-2 微結構 33 5-4-3 離子交換液相色層分析 33 5-4-4 降解性 34 5-4-5 與CBD-RGD融合蛋白質的作用 34 第六章 結論 36 第七章 參考文獻 37 圖次 47 表次 71 附錄一、培養基之配置 73 附錄二、試劑與緩衝溶液之製備 74zh_TW
dc.language.isoen_USzh_TW
dc.publisher獸醫微生物學研究所zh_TW
dc.subjectgelatin nanoparticlesen_US
dc.subject明膠奈米顆粒zh_TW
dc.subjectglutaraldehydeen_US
dc.subjectpH valueen_US
dc.subjectparticle sizeen_US
dc.subjection exchange liquid chromatographyen_US
dc.subjectCBD-RGD fusion proteinen_US
dc.subject戊二醛zh_TW
dc.subjectpH值zh_TW
dc.subject粒徑zh_TW
dc.subject離子交換液相層析zh_TW
dc.subjectCBD-RGD融合蛋白質zh_TW
dc.title明膠奈米顆粒製備與特性研究zh_TW
dc.titlePreparation and Characterization of Gelatin Nanoparticlesen_US
dc.typeThesis and Dissertationzh_TW
item.cerifentitytypePublications-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.languageiso639-1en_US-
item.fulltextno fulltext-
item.grantfulltextnone-
item.openairetypeThesis and Dissertation-
Appears in Collections:微生物暨公共衛生學研究所
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