臺灣博碩士論文加值系統

疾病能夠通過疫苗來控制。因此，篩選高效且專一性的免疫原用於疾病預防及治療非常重要，同時適當地評估疫苗的效能在疫苗的開發上扮演相當重要的角色。傳統的疫苗研發方式曠日廢時，已經無法面對新興疾病層出不窮的挑戰。隨著蛋白質體學的發展，使得快速篩檢出高效的免疫原變成可能。因此，應用蛋白質體學的研究方法如微陣列技術建立疫苗篩選及評估平台是可行的策略。本論文使用唾液酸多醣（Sialyl-Tn，STn）為抗原進行小鼠的免疫刺激，經蛋白質晶片技術進行抗體反應的測試。含有KLH的STn醣類抗原，確實能在動物體內誘導出免疫反應，產生IgM及IgG的變化。其中，官能基改變的抗原，可以得到較佳的免疫誘導反應，是為高效抗原。所以蛋白質微陣列可作為評估疫苗效能的參考。此外，我們也以市售流感疫苗進行相同的篩檢，得知流感疫苗的確會使人體的IgG效價持續上升，證實該平台亦可應用於臨床疫苗效價評估。
本論文主要目的在建立各式疫苗之效益評估及篩選的平台模式，於未來可提供疫苗研發之應用。

Vaccine was an excellent strategy of disease control. Screening of high efficiency and specificity immunogenic agents and evaluation of efficiency after vaccine injection are both important for vaccine research and development. Due to time consuming of vaccine research and development, the traditional techniques was not suitable for the condition of current pathogen changes.The issue of quickly and high-throughput screening for vaccine candidate was breakthrough after the proteomics technology, such as protein array, development.After the sailyl-Tn(STn) triggered the mice immune response, we used the microarray to detect the antibody reactivity, and found the IgM and IgG elevated in vivo. At the same way, we also found the high effectiveness vaccine agent was one of the epitope modified by specific functional group. We also using the common flu vaccine undergoing the same procedure, the efficiency were also evaluated. The protein array technique was an excellent tool for both preclinical and clinical evaluation for vaccine effectiveness. These finding was provided efficiency tool for vaccine research and development.

中文摘要 1
英文摘要 2
目錄 3
圖表目錄 5
第一章 緒論 8
1.1 疫苗與疾病 8
1.2 癌症與免疫治療 11
1.2.1 碳水化合物在細胞表面上扮演極重要的角色 15
1.2.2 聚醣的生物學特點 19
1.2.3 腫瘤相關性聚醣的結構改變 20
1.2.4 碳水化合物與腫瘤疫苗 23
1.3 腫瘤疫苗的作用 29
1.3.1 腫瘤疫苗的種類及其作用原理 29
1.3.2 疫苗的有效性評估 40
1.4 蛋白質晶片 41
1.4.2 微流體晶片技術 43
第二章 實驗目的 49
第三章 研究材料與方法 51
3.1 實驗材料 51
3.1.1 化學藥品 51
3.1.2 試藥成份 52
3.1.3 抗體 53
3.2 實驗方法 55
3.2.1 動物實驗 55
3.2.2 蛋白質晶片製作 57
3.2.3 疫苗效價的評估 58
3.2.4 西方墨點分析法 (Western blot analysis)： 59
3.2.5 單因子變異數分析(One-Way ANOVA)統計 59
第四章 實驗結果與分析 61
4.1 疫苗抗原的取得 61
4.2 動物免疫注射 61
4.3 蛋白質晶片製作 61
4.4 合成抗原疫苗效能的測試 62
4.5 流感疫苗效能的測試 64
第五章 討論 66
5.1 疫苗的選擇 66
5.2 血清效價的評估 66
5.3 蛋白質晶片的製作與使用 67
5.4 合成STN與癌細胞STN的探討 68
5.5 流感疫苗的結果 69
5.6 快速篩選平台的應用 70
第六章 結論 71
附錄 73
參考文獻 110

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