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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38834
標題: | 毛細液相層析與拋棄式微電灑探針電灑法質譜/質譜於電白質鑑定之開發與應用 |
作者: | Chu-Yun Tsai 蔡竺芸 |
指導教授: | 何國榮 |
關鍵字: | 質譜,蛋白質, mass spectrometry,protein identification, |
出版年 : | 2005 |
學位: | 碩士 |
摘要: | 我們以簡單的設備建立毛細液相層析電灑法串聯質譜儀,成功地鑑定大白鼠第二型肺泡細胞特有胞器-層質體 (lamellar bodies) 之組成蛋白質,有助於未來臨床上對呼吸窘迫症病患的處理與治療。有鑑於層析方法過於耗時,本研究第二部分為開發簡單且可拋棄式的灑探針,將消化並淨化後的胜肽直接送入串聯質譜儀進行分析,可達到快速鑑定蛋白質及無樣品殘留之問題;以肌紅蛋白 (Myoglobin) 的胰蛋白酶 (trypsin) 消化胜肽為樣品,蛋白質身分鑑定的極限可至2.5 fmol,具有使用方便及低樣品消耗的優點。第三部分我們以濃縮為主訴求,希望能開發簡單、實用且能線上淨化的方法,所開發之線上淨化微電灑串聯質譜儀,可大幅提高濃縮倍率及減少分析時間,實際應用於層質體的蛋白質體,得到高可信度的蛋白質鑑定結果。 We set up an on-line micro capillary reversed phase liquid chromatography interfaced to a tandem mass spectrometry (μLC-MS/MS) for protein identification. Proteins from lamellar bodies which were separated by two-dimensional gel electrophoresis were identified by μLC-MS/MS. However, the current technique is rather time consuming. To reduce the analysis time, we developed a disposable device that combined on-line clean up and infusion unseparated proteolytic peptides into an ESI tandem MS. It was demonstrated that several proteins of lamellar bodies could be successfully identified by using this on-line clean up method. Compared to μLC-MS/MS, this method could identify proteins rapidly and without the problem of cross-contamination although the peptide sequence coverage was slightly lower. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38834 |
全文授權: | 有償授權 |
顯示於系所單位: | 化學系 |
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