Iron(lll)-immobilized metal ion affinity chromatography and mass spectrometry for the purification and characterization of synthetic phosphopeptides

被引：96

作者：

Li, SH ^{[1
]}

Dass, C ^{[1
]}

机构：

[1] Univ Memphis, Dept Chem, Memphis, TN 38152 USA

来源：

ANALYTICAL BIOCHEMISTRY | 1999年 / 270卷 / 01期

关键词：

D O I：

10.1006/abio.1999.4060

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method based upon immobilized metal ion affinity chromatography (IMAC) is described for purification of phosphopeptides from the crude preparations of solid-phase peptide synthesis step. Affinity chromatography consists of iron(III) immobilized on iminodiacetate-agarose gel. The method was applied for purification of seven synthetic enkephalin-related phosphorylated peptides. The effectiveness of the method was evaluated by analyzing the IMAC-retained and -nonretained components using reversed-phase (RP) high-performance liquid chromatography (HPLC) and an on-line combination of RP-HPLC and electrospray ionization mass spectrometry. The UV and total ion current chromatograms demonstrated that the phosphopeptides were effectively separated and purified. (C) 1999 Academic Press.

引用

页码：9 / 14

页数：6

共 18 条

[1] ISOLATION OF PHOSPHOPROTEINS BY IMMOBILIZED METAL (FE-3+) AFFINITY-CHROMATOGRAPHY [J].