Automated 2D Peptide Separation on a 1D Nano-LC-MS System

被引：55

作者：

Taylor, Paul ^{[1
]}

Nielsen, Peter A. ^{[3
]}

Trelle, Morten B. ^{[3
]}

Horning, Ole B. ^{[3
]}

Andersen, Michael B. ^{[3
]}

Vorm, Ole ^{[3
]}

Moran, Michael F. ^{[1
,2
,4
]}

Kislinger, Thomas ^{[5
]}

机构：

[1] Hosp Sick Children, Program Mol Struct & Funct, Toronto, ON M5G 1X8, Canada

[2] Univ Toronto, Dept Mol Genet, Toronto, ON M5S 1A8, Canada

[3] Proxeon Biosyst, Odense, Denmark

[4] Univ Toronto, Dept Med Biophys, McLaughlin Ctr Mol Med, Toronto, ON M5G 1L7, Canada

[5] Ontario Canc Inst, Toronto, ON M5G 2M9, Canada

来源：

JOURNAL OF PROTEOME RESEARCH | 2009年 / 8卷 / 03期

基金：

加拿大健康研究院;

关键词：

Proteomics; MudPIT; nano-LC; 2D peptide separation; vented column; PROTEIN IDENTIFICATION TECHNOLOGY; MASS-SPECTROMETRY; SHOTGUN PROTEOMICS; STATISTICAL-MODEL; ELECTROPHORESIS; FRACTIONATION; MAMMALS; COLUMN; MOUSE; SERUM;

D O I：

10.1021/pr800986c

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Given the complexity of the mammalian proteome, high-resolution separation technologies are required to achieve comprehensive proteome coverage and to enhance the detection of low-abundance proteins. Among several technologies, Multidimensional Protein Identification Technology (MudPIT) enables the on-line separation of highly complex peptide mixtures directly coupled with mass spectrometry-based identification. Here, we present a variation of the traditional MudPIT protocol, combining highly sensitive chromatography using a nanoflow liquid chromatography system (nano-LC) with a two-dimensional precolumn in a vented column setup. When compared to the traditional MudPIT approach, this nanoflow variation demonstrated better first-phase separation leading to more proteins being characterized while using rather simple instrumentation and a protocol that requires less time and very little technical expertise to perform.

引用

页码：1610 / 1616

页数：7

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