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研究生: 張承俊
Cheng-Chun Chang
論文名稱: 奈米鑽石結合質譜技術分析醣蛋白
The Application of Nanodiamond and Mass Spectrometry in Glycoproteins Analysis
指導教授: 張煥正
Chang, Huan-Cheng
學位類別: 碩士
Master
系所名稱: 化學系
Department of Chemistry
論文出版年: 2009
畢業學年度: 97
語文別: 中文
論文頁數: 58
中文關鍵詞: 奈米鑽石質譜
論文種類: 學術論文
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  • 我們發展了一套方法,此方法可以加快、簡化在分析醣蛋白上中性醣醣基時的實驗流程,而且同時也可以增強在利用基質輔助雷射脫附游離飛行時間質譜儀分析醣類時醣類的訊號強度。
      此方法包括了利用微波加熱輔助進行醣蛋白的酵素消化及切醣,接著利用酸洗過後含有羧基的奈米鑽石進行純化,將胜肽移除,最後將待測樣品溶液先與氫氧化鈉溶液混合,再與基質(DHB)混合,以抑制在質譜儀分析時未移除乾淨的胜肽及醣類接鉀([M+K]+)的離子訊號。
      利用此方法所展現的優點,皆是我們使用基質輔助雷射脫附游離飛行時間質譜儀分析Ovalbumin和Ribonuclease B這兩種醣蛋白上所切下的N-linked的醣類當作樣品所得到的結果。

    Abstract 3 論文摘要 4 Chapter 1 5 緒論 5 1.1 奈米鑽石 5 1.2 醣蛋白及醣 13 Chapter 2 22 實驗部份 22 2.1 實驗藥品 22 2.2 實驗儀器 24 2.3  實驗步驟 26 a.酸洗奈米鑽石: 26 b.測試酸洗後的奈米鑽石會不會吸附醣類: 26 c.利用傳統加熱方法做醣蛋白的消化及切醣: 27 d.改良微波反應器加熱管: 27 e.測試Trypsin digest微波加熱反應的條件: 28 f.測試PNGase F切醣微波加熱反應的條件: 29 g.利用微波加熱方法做醣蛋白的消化及切醣: 29 h.利用酸洗過後的奈米鑽石做醣的純化: 30 i.MALDI樣品的製備: 30 Chapter 3 33 結果與討論 33 3.1 測試酸洗後的奈米鑽石會不會吸附醣類: 33 3.2 利用傳統加熱方法做醣蛋白的消化及切醣加上奈米鑽石分離及質譜分析: 36 3.3 測試微波反應的條件: 37 3.4 利用微波加熱方法做醣蛋白的消化及切醣加上奈米鑽石分離及質譜分析: 40 3.5 改善基質條件將醣類接鉀的離子訊號被抑制掉: 41 3.6 改善基質條件降低胜肽的偵測靈敏度: 47 3.7 結合奈米鑽石純化及改善基質進行醣蛋白分析: 48 a.傳統加熱方法: 48 b.微波加熱方法: 50 3.8 分析醣基為酸性醣的醣蛋白: 53 Chapter 4 56 結論 56 參考文獻 58

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