臺灣博碩士論文加值系統

本研究已經成功開發出結合液體依序釋放及磁鐵模組的自動化離心式蛋白質純化平台。這個平台在純化前先將所有試劑預先注入儲存槽中，利用馬達操作來控制試劑依照純化流程釋放，並利用磁鐵模組在捕捉、清洗及洗脫步驟時讓磁珠在反應槽中與液體充分混合，且排空液體時能將磁珠保留在反應槽中，改善傳統蛋白質純化費工的步驟使之自動化，整個純化步驟在1小時內就可以完成，與微量離心管內的蛋白質純化做比較亦不遜色。

This study has successfully developed an automated protein purification approach on a centrifugal platform. The major techniques used on this platform are the liquid sequential flow control and the magnet module. During operation, all reagents were preloaded into the storage chambers. The sequential release of the reagents was conducted through rapid deceleration of the disk and the mixing, and washing of the magnetic beads were performed by the magnetic module. Finally, an automated protein purification platform, which is able to complete the purification process within 1 hour, was developed. It showed similar performance comparing to the approach conducted using the Eppendorf tubes.

第一章 緒論 9
1-1 蛋白質純化的應用 9
1-2 離心式微流體技術 9
1-3 研究動機 10
第二章 文獻探討 11
2-1 傳統蛋白質純化方法 11
2-2 微流體蛋白質純化方法 14
2-3 離心式微流體蛋白質純化技術 17
第三章 研究設計 18
3-1 實驗材料 18
3-1-1 蛋白質純化磁珠試劑 18
3-1-2 蛋白質檢測試劑 18
3-1-3 微流體碟片材料 18
3-1-4 磁鐵材料 18
3-1-5 實驗儀器 18
3-2 碟片製作 20
3-2-1 蛋白質純化碟片製作 20
3-2-2 磁鐵模組建立 22
3-3 實驗樣本製備 24
3-3-1 細菌培養 24
3-3-2 樣品製備 25
3-4傳統操作流程 26
3-4-1 磁珠預處理 26
3-4-2 目標蛋白與磁珠鍵結 26
3-4-3 清洗 26
3-4-4 洗脫 26
3-4-5 磁珠後處理 27
3-5 碟片操作流程 28
3-5-1 試劑注入及捕捉步驟 28
3-5-2 清洗步驟 28
3-5-3 洗脫及定量步驟 29
3-5-4 檢測步驟 29
3-6 分析檢測 30
3-6-1 馬達系統 30
3-6-2 高速攝影系統 31
3-6-3 照片分析系統 32
3-6-4 膠體電泳檢測系統 33
3-6-5 光學檢測系統 37
第四章 實證結果與分析 39
4-1 流體功能 40
4-1-1 液體傾注之原理 40
4-1-2 試劑依序釋放 41
4-1-3 最佳馬達操作條件 43
4-1-4 定量 46
4-2 磁鐵模組的結合 48
4-2-1 磁珠損失 48
4-2-2 磁鐵模組的設計 49
A. 移動磁鐵拖動磁珠 49
B. 固定磁鐵吸引磁珠 51
4-2-3清洗效率 52
第五章 結論 54
第六章 未來展望 55
參考文獻 56

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