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研究生:陳怡婷
研究生(外文):Yi-Ting Chen
論文名稱:琥珀酸酐化明膠微球的製備及其包覆陽離子蛋白質之研究
指導教授:顧野松
指導教授(外文):Ye-song Gu
學位類別:碩士
校院名稱:東海大學
系所名稱:化學工程學系
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
中文關鍵詞:明膠微球陽離子蛋白質交聯劑
外文關鍵詞:microsphereslysozymesuccinic anhydride
相關次數:
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明膠(Gelatin)是由動物皮骨萃取而得之生物可降解性材料,由於此特性使其於生醫方面有著相當廣泛的應用。明膠(Gelatin)極易溶於水且膨潤度高,故只能作為短效性之藥物載體。而經過化學交聯反應後則可控制明膠之膨潤度與降解速率,改善其短效性釋放的缺點。利用琥珀酸酐進行取代反應(pH>9)後,可使明膠之帶負電量增加,載體的極性亦增強,此點有助於與陽離子蛋白質藥物結合之能力,更有利於增加藥物包覆量。
本研究除了改善明膠性質外,亦將乳化機制的有機油相以植物油來替代,使乳化過程較為溫和無毒性。實驗結果顯示,琥珀酸酐化明膠(succinated gelatin;SG)無法單獨做為微球載體之材料,可添加明膠及介面活性劑促使其形成完整微球型態。再利用旋轉方式均勻混和(Rota-mix)微球與不同濃度之交聯劑進行交聯反應,以調控載體之膨潤性使其適合應用於藥物釋放調控系統。
體外釋放之實驗顯示,單位微球載體可包覆溶菌酶(lysozyme)量有其極大值。最初逐漸增加溶菌酶(lysozyme)添加量,單位微球包覆量也隨之增加,但當單位微球包覆量達飽和時,即使再增加溶菌酶(lysozyme)添加量,單位微球包覆量也不再改變。此外,溶菌酶(lysozyme)會隨著時間增加而逐漸由載體中釋出,且沒有突釋的現象,此結果則有達到藥物釋放調控系統的要求。
Gelatin is a kind of material with biological degradability, which is normally extracted from the animal skins and bones. This characteristic of biological degradability makes gelatin be widely used in the biomedical field. It can be easily dissolved in water with high swelling capacity; therefore it can only be used as a short term carrier for medicine. However, its swelling degree and degeneration speed can be controlled after chemical crosslinking process to overcome the shortcoming of short term release. Gelatin carries more numbers of negative charges after succinic anhydride substitution reaction (pH>9), this can increase the medicine loading through the enforced electrostatic interaction with positive charges protein, which is beneficial to increase the quantity of carried medicine.
Besides the improvement of gelatin's character, during this research we have also substituted the organic oil phase for emulsification with the plant oil phase. This will make the emulsion process more temperately and nontoxic. The experimental result shows that, succinated gelatin (SG) can not be used alone as the material for microspheres carrier, however, the microspheres can be formed by mixing SG with unreacted gelatin. Then we investigate the effect of different concentrations of crosslinking agents, such as glutaraldehyde(GA), or controlling the swelling capacity of carrier.
Releasing experiment in vitro demonstrates that there is a limitation for lysozyme loading per microspheres carrier. Below this limit value, the amount of lysozyme carried increases gradually with the increased amount of lysozyme. The lysozyme is gradually released from the carrier with time and there is no sudden release. This character enables it to achieve the requirements of medicine releasing system.
中文摘要 ------------------------------------------- Ⅰ
英文摘要 ------------------------------------------- Ⅲ
誌謝------------------------------------------------ Ⅴ
目錄 ----------------------------------------------- Ⅶ
表目錄 --------------------------------------------- Ⅹ
圖目錄 --------------------------------------------- XI


第一章 緒論 ---------------------------------------- 1
1-1 前言-------------------------------------------- 1
1-2 明膠簡介---------------------------------------- 4
1-3琥珀酸酐之取代----------------------------------- 11
1-4交聯反應----------------------------------------- 12
1-5 乳化聚合系統------------------------------------ 14
1-5.1 簡介------------------------------------------- 14
1-5.2乳化聚合系統----------------------------------- 16
1-6 溶菌酶(lysozyme)簡介----------------------------- 21
1-7 研究動機---------------------------------------- 28
第二章 實驗設備及材料------------------------------- 30
2-1 實驗藥品---------------------------------------- 30
2-2 實驗儀器設備------------------------------------ 33
第三章 實驗系統 ------------------------------------ 35
3-1 製備微球---------------------------------------- 35
3-1.1 明膠的取代反應--------------------------------- 35
3-1.2 乳化聚合法------------------------------------- 37
3-2 微球之交聯反應---------------------------------- 39
3-3 微球包覆蛋白質藥物------------------------------ 41
3-4 分析方法---------------------------------------- 44
3-4.1 蛋白質電泳分析(SDS-PAGE)------------------ 44
3-4.2 分光光度計(UV-vis)--------------------------- 50
3-4.3 蛋白質定量法----------------------------------- 52
3-4.4 場發掃描式電子顯微鏡(FE-SEM)及掃描式電子顯微鏡(SEM) -54
3-4.4.1 構造原理------------------------------------ 54
3-4.4.2 樣品處理------------------------------------ 55
3-4.5 Ninhydrin reaction--------------------------------56
第四章 實驗結果與討論 ------------------------------ 59
4-1 明膠微球之製備---------------------------------- 59
4-2 利用化學交聯法修飾微球載體----------------------- 68
4-3 包覆之蛋白質定量分析---------------------------- 75
4-4 界面活性劑濃度對包覆蛋白質的影響---------------- 79
4-5 明膠微球載體吸附蛋白質藥物---------------------- 81
第五章 結論與建議 ---------------------------------- 85
5-1 實驗結論---------------------------------------- 85
5-2 實驗建議---------------------------------------- 86
第六章 參考文獻 ------------------------------------ 88
附錄一---------------------------------------------- 97
簡歷------------------------------------------------ 98
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