Reversed-phase chromatography with multiple fraction concatenation strategy for proteome profiling of human MCF10A cells.
Reversed-phase chromatography with multiple fraction concatenation strategy for proteome profiling of human MCF10A cells.
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DOI：
10.1002/pmic.201000722
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发表时间：
2011-05
期刊：
影响因子：
3.4
通讯作者：
Smith, Richard D.
中科院分区：
文献类型：
--
作者：
Wang, Yuexi;Yang, Feng;Gritsenko, Marina A.;Wang, Yingchun;Clauss, Therese;Liu, Tao;Shen, Yufeng;Monroe, Matthew E.;Lopez-Ferrer, Daniel;Reno, Theresa;Moore, Ronald J.;Klemke, Richard L.;Camp, David G., II;Smith, Richard D.
在这项研究中，我们评估了一种串联的低pH(PH 3)和高pH(PH 10)反相色谱策略，作为对胰酶消化的人MCF10A细胞样本进行二维液相色谱串联质谱(“鸟枪”)蛋白质组学分析的第一维方法。与更传统的强阳离子交换方法相比，串联高pH反相液相色谱作为一维分离策略，使多肽和蛋白质的鉴定数量(具有两个或两个以上唯一多肽)分别增加了1.8倍和1.6倍。除了更广泛的鉴定外，串联高pH分级方法的优点还包括提高蛋白质序列覆盖率、简化样品处理和减少样品损失。结果表明，串联的高pH反相策略是一种有吸引力的替代强阳离子交换的二维蛋白质组学分析方法。
In this study, we evaluated a concatenated low pH (pH 3) and high pH (pH 10) reversed-phase liquid chromatography strategy as a first dimension for two-dimensional liquid chromatography tandem mass spectrometry (“shotgun”) proteomic analysis of trypsin-digested human MCF10A cell sample. Compared with the more traditional strong cation exchange method, the use of concatenated high pH reversed-phase liquid chromatography as a first-dimension fractionation strategy resulted in 1.8- and 1.6-fold increases in the number of peptide and protein identifications (with two or more unique peptides), respectively. In addition to broader identifications, advantages of the concatenated high pH fractionation approach include improved protein sequence coverage, simplified sample processing, and reduced sample losses. The results demonstrate that the concatenated high pH reversed-phased strategy is an attractive alternative to strong cation exchange for two-dimensional shotgun proteomic analysis.
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影响因子：
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