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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳秀男 | |
dc.contributor.author | Chang-Yu Tsai | en |
dc.contributor.author | 蔡昌佑 | zh_TW |
dc.date.accessioned | 2021-06-07T17:49:12Z | - |
dc.date.copyright | 2013-02-16 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-02-04 | |
dc.identifier.citation | 參考文獻
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/15637 | - |
dc.description.abstract | 本論文使用微膠囊包覆技術(Microencapsulation)包覆點帶石斑虹彩病毒(orange spotted grouper iridovirus, OSGIV)次單位疫苗,使用的包覆材質是褐藻酸鈉,具有無毒、不影響抗原的免疫性及低成本的優勢,以褐藻酸鈉當作包覆材質製成的微膠囊(Microcapsule)在酸性環境中相當穩定,不會被強酸崩解而釋出內含物。相對地,微膠囊在鹼性環境中會因為pH值改變而慢慢崩解,因此,微膠囊可以保護疫苗通過胃酸環境,抵達腸道,使疫苗在弱鹼環境的腸道內釋放。本論文使用幾丁聚醣作為添加物,增加微膠囊的蛋白質乘載率及包覆率,依據本論文的微膠囊製程法,蛋白質乘載率為7.74%±1.23%,包覆率為50.46%±8.03%。
將微膠囊以外添加的方式與魚用飼料混合,連續餵食4週。腸道內專一性抗體出現顯著差異的時間比血清中專一性抗體早。每週持續追蹤血清中專一性抗體變化,結果顯示,口服低劑量組的血清內專一性抗體足夠維持8週,口服高劑量組的血清內專一性抗體則維持10週。根據本論文研究的微膠囊製程法顯示,包覆點帶石斑虹彩病毒之次單位口服疫苗,可以成功將疫苗送至後腸並引發免疫反應,產生專一性抗體。 | zh_TW |
dc.description.abstract | The vacccine has to be protected against gastric acid and digestive system. My thesis uses microencapsulation technique to encapsulate OSGIV (orange spotted grouper iridovirus, OSGIV) subunit vaccine from digestive system. Microencapsulation material is sodium alginate, its properties are low toxic, low antigenic and low cost. Using sodium alginate as microencapsulation material is called microcapsule. It is acid-stable ,but it will be collapsed when pH value arises in base environment. Therefore, microcapsule can make vaccine safely passing through stomach and be released in postgut. In my thesis, microcapsule loading rate is 7.74%±1.23%,encapsulation efficiency is 50.46%±8.03%.
Mix microcapsule and grouper artificial feed and keep feeding Epinephelus lanceolatus for four weeks, I find out that the groups of fed oral vaccine’s specific antibody in gut arise faster then serum. The results are the group of fed low dosage oral vaccine’s specific antibody can keep eight weeks and the group of fed high dosage oral vaccine’s specific antibody can keep ten weeks. Based on my thesis, the microcapsule of oral vaccine can induce immune response to produce specific antibody successfully. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T17:49:12Z (GMT). No. of bitstreams: 1 ntu-102-R99b45008-1.pdf: 1389633 bytes, checksum: 2f48c862bff4757d5ccc112656d9a2f2 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 目錄
謝辭 I 中文摘要 II 英文摘要 III 目錄 IV 第一章 文獻整理 1 第一節 石斑魚簡介 1 第二節 虹彩病毒 2 2.1虹彩病毒的分類地位及特性 2 2.2虹彩病毒感染症狀 3 2.3 虹彩病毒防治策略 3 第三節 魚類疫苗種類 4 3.1 次單位疫苗 (Subunit Vaccine) 4 3.2 虹彩病毒抗原蛋白片段-錐體膜蛋白 6 第四節 魚用疫苗給予方法 6 4.1 注射法 6 4.2 浸泡法 7 4.3 口服法 7 第五節 口服疫苗的包覆 8 5.1 微膠囊化的包覆材質 9 5.2 褐藻膠 9 5.3 褐藻膠的凝膠化 10 5.4 幾丁聚醣 11 5.5 幾丁聚醣-褐藻膠的凝膠作用 12 第六節 實驗目的 12 第二章 實驗材料與方法 13 第一節 利用大腸桿菌蛋白質表現系統生產虹彩病毒錐體膜蛋白 13 1.1 製作大腸桿菌BL21勝任細胞 13 1.2 pGEX-MMP質體轉型至大腸桿菌BL21勝任細胞 13 1.3 聚合酶連鎖反應確認已轉型之大腸桿菌BL21勝任細胞 14 1.4 蛋白質表現技術表現重組蛋白 14 1.5 測定大腸桿菌提取液總蛋白含量 15 1.6 蛋白質膠體電泳分析大腸桿菌提取液 15 1.7 西方墨點法分析大腸桿菌提取液 15 第二節 製作微膠囊 16 2.1 褐藻酸鈉-氯化鈣包覆大腸桿菌提取液 16 2.2 鏡檢微膠囊型態 17 2.3西方墨點法(Western-blot)分析微膠囊內含物 17 2.4微膠囊特性常數分析 18 2.5測試褐藻酸鈉包覆蛋白於胃蛋白酶及腸道環境的穩定性 18 2.6將製成的微膠囊與人工飼料混合 19 第三節 龍膽石斑體內試驗專一性抗體分析 19 3.1 實驗動物 19 3.2 實驗設計 19 3.3分析龍膽石斑胃、腸道內pGEX-MMP-CM釋放結果 20 3.4測定龍膽石斑免疫組織內疫苗變化量 20 3.5測定龍膽石斑胃、腸道及血清內專一性抗體 21 第四節 統計分析 21 第三章 實驗結果 22 第一節 利用大腸桿菌蛋白質表現系統生產虹彩病毒錐體膜蛋白 22 1.1 重組質體(pGEX-MMP)轉型後之大腸桿菌BL21確認 22 1.2 OSGIV-8L 蛋白質序列之分析 22 1.3 OSGIV-8L重組蛋白之蛋白質產量分析 22 1.4 OSGIV-8L重組蛋白之大腸桿菌提取液分析 23 第二節 製作微膠囊 23 2.1微膠囊製備條件之水相溶液/油相溶液測試 23 2.2分析pGEX-MMP-CM及pGEX-MMP-CAM微膠囊特性參數 24 2.3分析pGEX-MMP-CAM在不同模擬環境中的蛋白累積釋放率 24 第三節 龍膽石斑體內試驗專一性抗體分析 25 3.1檢測龍膽石斑是否曾被OSGIV感染 25 3.2 分析微膠囊免疫魚體後胃及腸道內專一性抗體變化 25 3.3 分析微膠囊免疫魚體後血清中專一性抗體變化 26 第四章 實驗討論 27 圖目錄 Figure 1. 確認重組質體之1.5%洋菜膠電泳圖 33 Figure 2. 虹彩病毒 OSGIV-8L 之蛋白序列穿膜結構預測圖 34 Figure 3. 重組質體蛋白質電泳分析圖 35 Figure 4. 以西方墨點法分析帶有重組質體之大腸桿菌提取液結果圖 36 Figure 5. 無法形成微膠囊型態圖 37 Figure 6. 鏡檢已包覆重組蛋白之微膠囊 38 Figure 7. 將包覆重組蛋白之微膠囊以pH12 ddH2O處理後之鏡檢圖 39 Figure 8. 微膠囊特性參數分析之蛋白質乘載率 40 Figure 9. 微膠囊特性參數分析之微膠囊包覆率 41 Figure 10. 西方墨點法分析有無加入幾丁聚醣的組別之結果圖 42 Figure 11. 微膠囊於SIF及SGF累積蛋白釋放率結果圖 43 Figure 12. 確認實驗動物是否帶原之1.5%洋菜膠電泳圖 44 Figure 13. 胃、前腸及後腸之蛋白質內含物電泳分析之銀染圖 45 Figure 14. 測定餵食疫苗後組織內pGEX-MMP變化圖 46 Figure 15. 測定胃內含物之專一性抗體變化圖 47 Figure 16. 測定前腸內含物之專一性抗體變化圖 48 Figure 17. 測定後腸內含物之專一性抗體變化圖 49 Figure 18. 測定口服低劑量組腸道內含物之專一性抗體變化圖 50 Figure 19. 測定口服高劑量組腸道內含物之專一性抗體變化圖 51 Figure 20. 測定血清內專一性抗體變化圖 52 Figure 21. 微膠囊大小、攪拌速度及乳化劑(Span 80)之三相圖 (Rodrigues et al., 2006) 53 Figure 22. 褐藻酸鈉與幾丁聚醣形成聚電解膜之化學反應簡圖 (Xue et al., 2004) 54 表目錄 Table 1. 不同比例之油相溶液/水相溶液形成微膠囊一覽表 55 參考文獻 56 附錄 70 | |
dc.language.iso | zh-TW | |
dc.title | 點帶石斑虹彩病毒次單位口服疫苗及包覆技術之研發 | zh_TW |
dc.title | Development of orange-spotted grouper iridovirus (OSGIV) myristylated membrane protein subunit oral vaccine preparation and microencapsulation | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 王俊順,黃世鈴,冉繁華 | |
dc.subject.keyword | 口服疫苗,點帶石斑虹彩病毒,微膠囊,微膠囊包覆技術,褐藻酸鈉, | zh_TW |
dc.subject.keyword | oral vaccine,orange spotted grouper iridovirus,microencapsulation,microcapsule,sodium alginate, | en |
dc.relation.page | 76 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2013-02-04 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
顯示於系所單位: | 漁業科學研究所 |
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