Development of Efficient Protein Extraction Methods for Shotgun Proteome Analysis of Formalin-Fixed Tissues

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• 作者：Xiaogang Jiang ; Xinning Jiang ; Shun Feng ; Ruijun Tian ; Mingliang Ye ; Hanfa Zou
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：March 2007
• 年：2007
• 卷：6
• 期：3
• 页码：1038 - 1047
• 全文大小：354K
• 年卷期：v.6,no.3(March 2007)
• ISSN：1535-3907

文摘

There are vast archives of formalin-fixed tissues spanning many conceivable conditions such as differentdiseases, time courses, and different treatment and allowing acquisition of the necessary numbers ofsamples to carry out biomarker discovery study. However, the conventional protein analysis approachis not applicable for the analysis of proteins in the formalin-fixed tissue because the formalin fixationprocess resulted in the cross-linking of proteins, and thus, intact proteins cannot be efficiently extracted.In this study, several protocols were investigated to extract proteins from formalin-fixed mouse livertissue for shotgun proteome analysis. It was found that incubation of tissue in a lysis buffer containing6 M guanidine hydrochloride at high temperature led to the highest protein yield and the largest numberof proteins identified. The peptides and proteins identified from formalin-fixed tissue were firstcomprehensively compared with those identified from frozen-fresh tissue. It was found that a majorityof peptides identified from fixed tissue were unmodified and proteome coverage for the analysis offixed tissue was not obviously compromised by the formalin fixation process. Valuable proteomeinformation could be obtained by shotgun proteome analysis of formalin-fixed tissue, which presentsa new approach for disease biomarker discovery.