One-Step Procedure for Peptide Extraction from In-Gel Digestion Sample for Mass Spectrometric Analysis

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• 作者：Wei Meng ; Huoming Zhang ; Tiannan Guo ; Chhiti Pandey ; Yi Zhu ; Oi Lian Kon ; Siu Kwan Sze
• 刊名：Analytical Chemistry
• 出版年：2008
• 出版时间：December 15, 2008
• 年：2008
• 卷：80
• 期：24
• 页码：9797-9805
• 全文大小：189K
• 年卷期：v.80,no.24(December 15, 2008)
• ISSN：1520-6882

文摘

Protein identification from samples resolved by one-dimensional and two-dimensional gel electrophoresis is highly dependent on the recovery of trypsin-digested peptides prior to mass spectrometric analysis. The commonly used two-step protocol for extracting tryptic peptides, involving high-volume organic solvent extraction and cleanup via microscale reversed-phase micropipet tip or microcolumn is not only limited by significant sample loss but is also costly and very labor-intensive. We report here a simple one-step procedure for simultaneous peptide extraction and cleanup by incubating a small piece of C18 Empore Disk (3M) with the in-gel digested solution. We show that the direct Empore Disk-based peptide extraction procedure is convenient, economical, and has higher efficiency as compared with the commonly used two-step protocol for peptide preparation prior to MS analysis.