色谱

• 研究论文 • 上一篇    下一篇

强阳离子交换色谱分离多肽混合物的条件优化

王璐1, 周兰兰1, 钱小红2, 张养军2*   

  1. 1. 安徽医科大学, 安徽 合肥 230032; 2. 蛋白质组学国家重点实验室, 军事医学科学院放射与辐射医学研究所, 北京蛋白质组研究中心, 北京 102206
  • 收稿日期:2009-10-12 修回日期:2009-12-12 出版日期:2010-04-28 发布日期:1980-12-25
  • 通讯作者: 张养军

Optimization of conditions for the separation of peptide mixtures using strong cation exchange chromatography

WANG Lu1, ZHOU Lanlan1, QIAN Xiaohong2, ZHANG Yangjun2*   

  1. 1. Anhui Medical University, Hefei 230032, China; 2. State Key Laboratory of Proteomics, Beijing Institute of Radiation Medicine, Beijing Proteome Research Center, Beijing 102206, China
  • Received:2009-10-12 Revised:2009-12-12 Online:2010-04-28 Published:1980-12-25
  • Contact: ZHANG Yangjun

摘要: 以酵母全蛋白提取液的胰蛋白酶切产物为研究对象,对多维色谱分离中常用的强阳离子交换色谱的分离条件,包括上样量、盐的种类、调节缓冲液pH值的酸的种类及有机溶剂的比例进行了考察和比较。结果表明,在强阳离子交换色谱分离线性上样量范围内,在氯化铵溶液作为流动相,用磷酸调节流动相缓冲液pH值至2.7,且流动相中乙腈的体积分数为30%条件下进行梯度洗脱时,可获得最佳的分离结果。本结果可为采用二维色谱-质谱策略分析多肽混合物时的第一维强阳离子交换色谱分离条件的选择提供有益的参考。

关键词: 蛋白质组学, 多肽分离 , 强阳离子交换色谱

Abstract: The separation conditions for strong cation exchange chromatography were investigated and compared using tryptic digest of protein extract from yeast. The conditions included the loading amount of the sample, the type of the salt in mobile phase, the type of the acid used to adjust the pH of the mobile phase and the proportion of organic solvent added in the elution solution. The experimental results indicated that high separation efficiency can be obtained by using ammonium chloride solution as optimal mobile phase, phosphoric acid to adjust the pH of mobile phases and 30%(v/v) organic solvent in mobile phases. The results suggested that a beneficial reference was presented for the selection of separation conditions of strong cation exchange chromatography when applying in two-dimensional chromatography-mass spectrometry strategy in proteomic research.

Key words: peptide separation , proteomics, strong cation exchange chromatography