PICquant: a quantitative platform to measure differential peptide abundance using dual-isotopic labeling with 12C6- and 13C6-phenyl isocyanate.
PICquant: a quantitative platform to measure differential peptide abundance using dual-isotopic labeling with 12C6- and 13C6-phenyl isocyanate.
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PICquant：使用 12C6- 和 13C6- 异氰酸苯酯双同位素标记测量差异肽丰度的定量平台。
DOI：
10.1021/ac102461e
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发表时间：
2011
影响因子：
7.4
通讯作者：
Templeton,DennisJ
中科院分区：
文献类型：
--
作者：
Lyons,CharlesE;Victor,KenG;Moshnikov,SergeyA;Bachmann,LorinM;Baras,AlexanderS;Dettmann,KathleenM;Cross,JanetV;Templeton,DennisJ
我们已经开发了一个完整的系统，用于同位素标记，分馏和自动定量的差异表达的肽，显着促进候选生物标志物的发现。我们描述了一种新的稳定的质量标记试剂对，12 C6-和13 C6-苯基异氰酸酯（PIC），提供了显着的优势，目前可用的标签。肽主要在其氨基末端标记，并显示出不依赖于标记同位素的洗脱曲线。重要的是，PIC标记的肽在CID片段化时具有独特的中性质量损失，其使得能够进行电荷状态和标记同位素鉴定，从而将序列鉴定与候选生物标志物的定量分离。为了利用这些特性，我们将肽分级分离方案与Thermo LTQ-XL LC-MS 2数据采集策略和一套自动光谱分析软件相结合，该软件可识别标记样品之间的定量差异。这种被称为PICquant平台的方法不依赖于蛋白质序列鉴定，并且排除了未标记的肽，否则会混淆生物标志物的发现。将PICquant平台应用于一组复杂的临床样本表明，该系统可以快速识别对照组和患者组之间差异表达的肽。
We have developed a complete system for the isotopic labeling, fractionation, and automated quantification of differentially expressed peptides that significantly facilitates candidate biomarker discovery. We describe a new stable mass tagging reagent pair,12C6- and13C6-phenyl isocyanate (PIC), that offers significant advantages over currently available tags. Peptides are labeled predominantly at their amino termini and exhibit elution profiles that are independent of label isotope. Importantly, PIC-labeled peptides have unique neutral-mass losses upon CID fragmentation that enable charge state and label isotope identification and, thereby, decouple the sequence identification from the quantification of candidate biomarkers. To exploit these properties, we have coupled peptide fractionation protocols with a Thermo LTQ-XL LC-MS2data acquisition strategy and a suite of automated spectrum analysis software that identifies quantitative differences between labeled samples. This approach, dubbed the PICquant platform, is independent of protein sequence identification and excludes unlabeled peptides that otherwise confound biomarker discovery. Application of the PICquant platform to a set of complex clinical samples showed that the system allows rapid identification of peptides that are differentially expressed between control and patient groups.