LC/MS/MS配合膠體中消化方法之蛋白質體學分析應用

The Applications of Proteomic Analysis by LC/MS/MS and In-Gel Digestion Method

方嘉德(Jia-Der Fang) ； 林曉薇(Hsiao-Wei Lin) ； 林曉莉(Hsiao-Li Lin) ； 吳烘(Hung Wu) ； 吳定峰(Ting-Feng Wu)

《化學》 61卷2期 (2003/06) Pp. 277-284

https://doi.org/10.6623/chem.2003028

摘要

本研究方法嘗試建立LC/MS/MS配合溶液中消化方法或者膠體中消化方法等兩種蛋白質之胺基酸定序分析工作平台。其中，本研究方法所採用的游離源是ㄧ般的電灑方式，而不是常見的nanospray方式。在本研究方法中，完成層析分離之後，會再將層析質譜以TurboSEQUEST進行資料庫比對工作。由本研究方法所得到的比對結果，其Xcorr常數均可達到1.0以上，且deltCn常數也都在0.2到1.0之間。而所得到之比對結果其Protein Coverage數值，無論是溶液中消化方法或者膠體中消化方法所獲得的結果都可以達到20%以上。此外在本研究中，也比較了Coomassie blue染色方法與銀染法以及銀染之後再脫色等各種處理方式所獲得之結果的差異性。

關鍵字

LC/MS/MS方法 ； 溶液中消化方法 ； 膠體中消化方法

並列摘要

This paper has tried to develop two amino acid sequencing method of protein by LCMS/MS with in-solution digestion method or in-gel solution method. These methods didn't use the nanospray source but the traditional ESI source. When we have got the chromatogram, we took the TurboSEQUEST database search. About the results of TurboSEQUEST database search, the xcorr values were always above 1.0, and the constants of deltCn were between 0.2 and 1.0. Whatever in solution digestion method or in-gel digestion method, all of their Protein Coverage values were greater than 20%. We also have compared the results between Coomassie Brilliant Blue stained method, silver stained method and silver stained-and-destained method.

並列關鍵字

LC/MS/MS ； in-solution digestion ； in-gel digestion