一种基于定量驱动策略和18O标记方法的比较蛋白质组学分析平台

定量蛋白质组学技术是一种高通量的蛋白质分析技术, 有望大大推动生物学和医学研究的发展. 最近提出了一种基于定量驱动的定量蛋白质组学分析新策略, 它采用定量和定性分析分开的模式, 先通过液相色谱一级质谱(LC-MS)定量分析确定差异肽段, 然后针对这些差异肽段再做液相色谱串联质谱(LC-MS/MS)定性分析, 最后通过定性和定量数据的匹配来获得差异蛋白结果. 这种策略被证明可以针对性地鉴定差异肽段, 增加差异蛋白的检测个数. 我们基于该策略和18O标记方法建立了一种新的定量蛋白质组学分析平台. 研究证明了该分析平台在定量蛋白质组学研究中的可行性. 本文还根据此研究在理论上提出了一种新的MS/MS分析的数据依赖模式, 即实时定量依赖的MS/MS离子选择模式.

A platform for quantitative proteomics based on a quantification-driven strategy and the 18O labeling method

As a new technology for determining protein expression, quantitative proteomics has great potential for application. Recently, a quantification-driven strategy utilizing quantitative proteomics was reported. Based on a quantification and identification-decoupled approach, this strategy first quantified the peptides by liquid chromatography mass spectrometry to confirm the differential peptides, and then targeted these differential peptides to liquid chromatography mass spectrometry/mass spectrometry analysis for identification, and ultimately quantified proteins by correlating and matching the quantitative and qualitative data. This strategy has been shown to selectively identify differential proteins and increase the quantity of differential proteins. Based on this strategy and the 18O labeling method, we developed a new platform in quantitative proteomics and the present study demonstrated the feasibility of this platform. According to this work, a new data-dependent mode for ion selection in mass spectrometry/mass spectrometry analysis was presented and has been coined as a real-time quantification-dependent mode.