质谱智能选择反应监测用于蛋白质绝对定量中母子离子对确证的方法研究

质谱选择反应监测(SRM)技术在蛋白质绝对定量分析中的应用越来越广泛,其成功应用的关键是肽段母子离子对的正确选择与确证.触发采集二级图谱是目前常用的母子离子对确认方法,但分析效率有待于提高.质谱智能选择反应监测(iSRM)是新近发展的高通量分析方法.为了考察该方法用于通量化母子离子对确证时的效果,选用牛血清白蛋白(BSA)和酵母蛋白提取物作为样品进行分析.结果表明,该方法具有更高的分析灵敏度,可以对低至1 fmol BSA样品中的母子离子对进行确证,而且相对于触发采集二级图谱方法而言,具有更高的分析通量,为规模化母子离子对选择与确证提供了一种新的策略.

Validation of Transitions by Intelligent Selected Reaction MonitoringMass Spectrometry for Protein Absolute Quantitation

Selected reaction monitoring(SRM)-mass spectrometry is increasingly applied to absolute quantitation of proteins.In large scale quantitative analyses,one of the main challenges is that the initial set of transitions should be optimized and validated to ensure that the quantified signals indeed derive from the targeted peptide.SRM-triggered MS/MS acquisition is an efficient way for validation,but still needs improvement.Recently,an innovative intelligent SRM(Isrm) was introduced to alleviate the challenge.In this study,iSRM was tested to validate the transitions of peptides of bovine serum albumin(BSA) and yeast proteins.The experimental results show that the method has increased selectivity and throughput for validation of transitions of peptides.Even for 1 fmol BSA,a set of transitions can be successfully validated,indicating that iSRM method is suitable for large scale validation of transitions of peptides in quantitative proteomics research.