Phosphopeptide-selective column-switching RP-HPLC with a titania precolumn

被引：77

作者：

Kuroda, I ^{[1
]}

Shintani, Y ^{[1
]}

Motokawa, M ^{[1
]}

Abe, S ^{[1
]}

Furuno, M ^{[1
]}

机构：

[1] GL Sci Inc, Iruma, Saitama 3580032, Japan

来源：

ANALYTICAL SCIENCES | 2004年 / 20卷 / 09期

关键词：

D O I：

10.2116/analsci.20.1313

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A methodology of phosphopeptide-selective analysis coupled with column-switching HPLC utilizing titania as precolumn media is presented. Phosphopeptides were selectively enriched on titania packing within a protein/peptide mixture without any additional procedure, and analyzed by column-switching high-performance liquid chromatography. First, phospho-compounds were separated from complex mixtures by trapping them under acidic conditions on a titania packing, where non-phosphorylated compounds were effused out of the precolumn. Subsequently, phospho-compounds were desorbed from the titania column under a specific condition and analyzed. The behavior of phospho-compounds on a titania surface, especially adsorption/desorption, was precisely examined and optimized. A phosphoric buffer was successively employed for the elution of phosphopeptides on a titania surface by competition with the free phosphate group. From the successes of a selective concentration/analysis of phosphopeptides with column-switching HPLC with a titania precolumn, a novel phosphopeptide-selective RP-HPLC analysis has been shown to have an application possibility as a tool for phosphoproteomics.

引用

页码：1313 / 1319

页数：7

共 16 条

[1] Heterologous expression and catalytic properties of the C-terminal domain of starfish Cdc25 dual-specificity phosphatase, a cell cycle regulator [J].