Integrated Device for Online Sample Buffer Exchange, Protein Enrichment, and Digestion

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• 作者：Liangliang Sun ; Junfeng Ma ; Xiaoqiang Qiao ; Yu Liang ; Guijie Zhu ; Yichu Shan ; Zhen Liang ; Lihua Zhang ; Yukui Zhang
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：March 15, 2010
• 年：2010
• 卷：82
• 期：6
• 页码：2574-2579
• 全文大小：224K
• 年卷期：v.82,no.6(March 15, 2010)
• ISSN：1520-6882

文摘

An integrated sample treatment device, composed of a membrane interface and a monolithic hybrid silica based immobilized enzymatic reactor (IMER), was developed for the simultaneous sample buffer exchange, protein enrichment, and online digestion, by which for the sample buffer, the acetonitrile content was reduced to 1/10 of the initial one, and the pH value was adjusted from 3.0 to 8.0, compatible for online trypsin digestion. Furthermore, the signal intensity of myoglobin digests was improved by over 10 times. Such an integrated device was successfully applied to the online treatment of three protein eluates obtained by reverse-phase liquid chromatography (RPLC) separation, followed by further protein digest analysis with microreverse-phase liquid chromatography-electrospray ionization-tandem mass spectrometry (μRPLC-ESI-MS/MS). The experimental results showed that the performance of such an integrated sample treatment device was comparable to that of the traditional offline sample treatment method, including lyophilization and in-solution digestion. However, the consumed time was reduced to 1/192. All these results demonstrate that such an integrated sample treatment device could be further online coupled with protein separation, peptide separation, and identification, to achieve high-throughput proteome analysis.