Hao Piliang, Zhang Huoming, Sze Siu Kwan
School of Biological Sciences, Nanyang Technological University, Singapore, Singapore.
Methods Mol Biol. 2011;790:305-18. doi: 10.1007/978-1-61779-319-6_23.
In shotgun proteomics, peptide fractionation is essential for in-depth characterization of proteomes and the mapping of protein posttranslational modifications. Recently, a mix-mode chromatography [i.e., electrostatic repulsion hydrophilic interaction chromatography (ERLIC)] has been developed and found to be a versatile method in proteome characterization. Here, we use ERLIC to characterize the glycoproteome and phosphoproteome simultaneously. We also demonstrate that the ERLIC can be an alternative to the commonly used strong cation exchange chromatography for higher recovery of proteins during whole proteome analysis. These protocols can be easily adopted in most proteomics laboratories.
在鸟枪法蛋白质组学中，肽段分级分离对于蛋白质组的深入表征和蛋白质翻译后修饰的图谱绘制至关重要。最近，一种混合模式色谱法[即静电排斥亲水相互作用色谱法（ERLIC）]已被开发出来，并被发现是蛋白质组表征中的一种通用方法。在这里，我们使用ERLIC同时表征糖蛋白质组和磷酸蛋白质组。我们还证明，在全蛋白质组分析过程中，ERLIC可以替代常用的强阳离子交换色谱法，以实现更高的蛋白质回收率。这些方案在大多数蛋白质组学实验室中都可以轻松采用。
