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。
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.
出处
《色谱》
CAS
CSCD
北大核心
2010年第4期368-373,共6页
Chinese Journal of Chromatography
基金
国家重点基础研究规划项目(2006CB910803
2007CB914104)
国家自然科学基金项目(30621063
20635010
20735005
20875101)
蛋白质组学国家重点实验室项目(SKLP-K200807
SKLP-O200808)
关键词
强阳离子交换色谱
蛋白质组学
多肽分离
strong cation exchange chromatography
proteomics
peptide separation
