Using iRT, a normalized retention time for more targeted measurement of peptides.

Using iRT, a normalized retention time for more targeted measurement of peptides.
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使用 iRT(一种标准化保留时间)可以更有针对性地测量肽。

DOI:
10.1002/pmic.201100463
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发表时间:
2012-04
期刊:
影响因子:
3.4
通讯作者:
Rinner, Oliver
Rinner, Oliver
中科院分区:
生物学3区
文献类型:
--
作者:
Escher, Claudia;Reiter, Lukas;MacLean, Brendan;Ossola, Reto;Herzog, Franz;Chilton, John;MacCoss, Michael J.;Rinner, Oliver

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多反应监测(MRM)最近已成为利用质谱学对蛋白质进行有针对性的定量测量的选择方法。然而,该方法在一次运行中可以测量的多肽数量受到限制。如果知道每个肽的准确保留时间(RT),则可以通过计划采集来显著增加这个数字。在这里,我们提出了IRT,一个经验性推导的无量纲肽特异值,允许高度准确的RT预测。多肽的IRT是相对于一组标准的参考IRT多肽的固定数字,可以跨实验室和层析系统进行转移。我们表明,与电子RT预测相比,IRT有助于建立多路实验,其获取窗口比电子RT预测小4倍以上,从而提高了量化精度。IRTS可以由任何实验室确定并透明共享。IRT的概念已经在最广泛使用的MRM实验软件Skyline中实现。
Multiple reaction monitoring (MRM) has recently become the method of choice for targeted quantitative measurement of proteins using mass spectrometry. The method, however, is limited in the number of peptides that can be measured in one run. This number can be markedly increased by scheduling the acquisition if the accurate retention time (RT) of each peptide is known. Here we present iRT, an empirically derived dimensionless peptide-specific value that allows for highly accurate RT prediction. The iRT of a peptide is a fixed number relative to a standard set of reference iRT-peptides that can be transferred across laboratories and chromatographic systems. We show that iRT facilitates the setup of multiplexed experiments with acquisition windows more than 4 times smaller compared to in silico RT predictions resulting in improved quantification accuracy. iRTs can be determined by any laboratory and shared transparently. The iRT concept has been implemented in Skyline, the most widely used software for MRM experiments.
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