On-line Digestion System for Protein Characterization and Proteome Analysis

被引：45

作者：

Lopez-Ferrer, Daniel ^{[1
]}

Petritis, Konstantinos ^{[1
]}

Lourette, Natacha M. ^{[1
]}

Clowers, Brian ^{[1
]}

Hixson, Kim K. ^{[2
]}

Heibeck, Tyler ^{[1
]}

Prior, David C. ^{[1
]}

Pasa-Tolic, Ljiljana ^{[2
]}

Camp, David G., II ^{[1
]}

Belov, Mikhail E. ^{[1
]}

Smith, Richard D. ^{[1
]}

机构：

[1] Pacific NW Natl Lab, Div Biol Sci, Richland, WA 99352 USA

[2] Pacific NW Natl Lab, Environm Mol Sci Lab, Richland, WA 99352 USA

来源：

ANALYTICAL CHEMISTRY | 2008年 / 80卷 / 23期

关键词：

D O I：

10.1021/ac800927v

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

An efficient on-line digestion system that reduces the number of sample manipulation steps has been demonstrated for high-throughput proteomics. By incorporating a pressurized sample loop into a liquid chromatography-based separation system, both sample and enzyme (e.g., trypsin) can be simultaneously introduced to produce a complete, yet rapid digestion. Both standard proteins and a complex Shewanella oneidensis global protein extract were digested and analyzed using the automated online pressurized digestion system coupled to an ion mobility time-of-flight mass spectrometer, an ion trap mass spectrometer, or both. The system denatured, digested, and separated product peptides in a manner of minutes, making it amenable to on-line high-throughput applications. In addition to simplifying and expediting sample processing, the system was easy to implement and no cross-contamination was observed among samples. As a result, the online digestion system offers a powerful approach for high-throughput screening of proteins that could prove valuable in biochemical research (rapid screening of protein-based drugs).

引用

页码：8930 / 8936

页数：7

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