Automated immobilized metal affinity chromatography/nano-liquid chromatography/electrospray ionization mass spectrometry platform for profiling protein phosphorylation sites

被引：77

作者：

Ficarro, SB ^{[1
]}

Salomon, AR ^{[1
]}

Brill, LM ^{[1
]}

Mason, DE ^{[1
]}

Stettler-Gill, M ^{[1
]}

Brock, A ^{[1
]}

Peters, EC ^{[1
]}

机构：

[1] Novartis Res Fdn, Genom Inst, San Diego, CA 92121 USA

来源：

RAPID COMMUNICATIONS IN MASS SPECTROMETRY | 2005年 / 19卷 / 01期

关键词：

D O I：

10.1002/rcm.1746

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A versatile integrated system has been developed for the automated enrichment and analysis of phosphopeptides by immobilized metal affinity chromatography/nano-liquid chromatography/electrospray ionization mass spectrometry (IMAC/nano-LC/ESI-MS). This system utilizes two independently controlled high-performance liquid chromatography (HPLC) pumps, an autosampler and microvalves to prepare and elute samples into an ion trap mass spectrometer. The use of robust reversed-phase HPLC columns with integrated ESI emitter tips enables the reproducible detection and identification of low-femtomole quantities of phosphopeptides. The entire system is coordinated through a simple user interface by customized software. The ruggedness of the system is demonstrated by highly reproducible analyses of single and multi-protein digests, while its utility is demonstrated by the thorough evaluation of the relative immunoprecipitation efficiencies of several commercially available anti-phosphotyrosine antibodies. Copyright (C) 2004 John Wiley Sons, Ltd.

引用

页码：57 / 71

页数：15

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