• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国卫生研究院文献>ACS AuthorChoice >Application of Displacement Chromatography to OnlineTwo-Dimensional Liquid Chromatography Coupled to Tandem Mass SpectrometryImproves Peptide Separation Efficiency and Detectability for the Analysisof Complex Proteomes
【2h】

Application of Displacement Chromatography to OnlineTwo-Dimensional Liquid Chromatography Coupled to Tandem Mass SpectrometryImproves Peptide Separation Efficiency and Detectability for the Analysisof Complex Proteomes

机译：置换色谱法在线应用二维液相色谱-串联质谱提高了肽的分离效率和分析的可检测性复杂蛋白质组学

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The complexity of mammalian proteomes is a challenge in bottom-up proteomics. For a comprehensive proteome analysis, multidimensional separation strategies are necessary. Online two-dimensional liquid chromatography–tandem mass spectrometry (2D-LC-MS/MS) combining strong cation exchange (SCX) in the first dimension with reversed-phase (RP) chromatography in the second dimension provides a powerful approach to analyze complex proteomes. Although the combination of SCX with RP chromatography provides a good orthogonality, only a moderate separation is achieved in the first dimension for peptides with two (+2) or three (+3) positive charges. The aim of this study was to improve the performance of online SCX-RP-MS/MS by applying displacement chromatography to the first separation dimension. Compared to gradient chromatography mode (GCM), displacement chromatography mode (DCM) was expected to improve the separation of +2-peptides and +3-peptides, thus reducing complexity and increasing ionization and detectability. The results show that DCM provided a separation of +2-peptides and +3-peptidesin remarkably sharp zones with a low degree of coelution, thus providingfractions with significantly higher purities compared to GCM. In particular,+2-peptides were separated over several fractions, which was not possibleto achieve in GCM. The better separation in DCM resulted in a higherreproducibility and significantly higher identification rates forboth peptides and proteins including a 2.6-fold increase for +2-peptides.The higher number of identified peptides in DCM resulted in significantlyhigher protein sequence coverages and a considerably higher numberof unique peptides per protein. Compared to conventionally used salt-basedGCM, DCM increased the performance of online SCX-RP-MS/MS and enabledcomprehensive proteome profiling in the low microgram range.
机译：哺乳动物蛋白质组的复杂性是自下而上蛋白质组学的一个挑战。对于全面的蛋白质组分析，多维分离策略是必要的。在线二维液相色谱-串联质谱（2D-LC-MS / MS）将第一维的强阳离子交换（SCX）与第二维的反相（RP）色谱相结合，提供了一种分析复杂蛋白质组的有效方法。尽管SCX与RP色谱法的结合提供了良好的正交性，但是对于带有两个（+2）或三个（+3）正电荷的肽，在第一维中仅实现了适度的分离。这项研究的目的是通过将置换色谱法应用于第一分离尺寸来提高在线SCX-RP-MS / MS的性能。与梯度色谱模式（GCM）相比，置换色谱模式（DCM）有望改善+ 2-肽和+ 3-肽的分离，从而降低复杂性并增加电离和可检测性。结果表明，DCM提供了+2肽和+3肽的分离在具有低共洗脱度的非常尖锐的区域中，从而提供与GCM相比纯度更高的馏分。特别是，+ 2-肽被分成几个部分，这是不可能的在GCM中实现。在DCM中更好的分离导致更高的分离度的重现性和较高的识别率肽和蛋白质都包括+ 2-肽的2.6倍增加。DCM中鉴定出的肽数量较高，导致更高的蛋白质序列覆盖率和更高的数量每个蛋白质独特的肽的数量。与常规使用的盐基相比GCM，DCM提高了在线SCX-RP-MS / MS的性能，并启用了低微克范围内的综合蛋白质组分析。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号