Iron(lll)-immobilized metal ion affinity chromatography and mass spectrometry for the purification and characterization of synthetic phosphopeptides
Iron(lll)-immobilized metal ion affinity chromatography and mass spectrometry for the purification and characterization of synthetic phosphopeptides
复制标题
DOI：
10.1006/abio.1999.4060
复制
发表时间：
1999-05-15
影响因子：
2.9
通讯作者：
Dass, C
中科院分区：
文献类型：
--
作者：
Li, SH;Dass, C
描述了一种基于固定化金属离子亲和层析（IMAC）的方法，用于从固相肽合成步骤的粗制剂中纯化磷酸肽。亲和层析由固定在亚氨基二乙酸盐-琼脂糖凝胶上的铁(III)组成。该方法用于纯化七种合成的脑啡肽相关磷酸化肽。通过使用反相 (RP) 高效液相色谱 (HPLC) 以及 RP-HPLC 和电喷雾电离质谱的在线组合分析 IMAC 保留和非保留组分，评估了该方法的有效性。紫外和总离子流色谱图表明磷酸肽得到了有效的分离和纯化。 (C) 1999 年学术出版社。
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.