Statistical inference from multiple iTRAQ experiments without using common reference standards.

Statistical inference from multiple iTRAQ experiments without using common reference standards.
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DOI:
10.1021/pr300624g
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发表时间:
2013-02-01
影响因子:
4.4
通讯作者:
Ruczinski I
Ruczinski I
中科院分区:
生物学2区
文献类型:
--
作者:
Herbrich SM;Cole RN;West KP Jr;Schulze K;Yager JD;Groopman JD;Christian P;Wu L;O'Meally RN;May DH;McIntosh MW;Ruczinski I

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相对和绝对定量等压标签(ITRAQ)是一种重要的蛋白质鉴定和定量的质谱学技术,能够在一次实验中分析多个样品。通常,iTRAQ实验使用其中一个通道中所有样品池中的等分或“主池”作为参考样品标准,以估计生物样品中蛋白质的相对丰度,并结合多个实验的丰度估计。在这份手稿中,我们表明使用主池是适得其反的。我们通过使用现有的生物数据而不是主池来获得对蛋白质相对丰度的更准确的估计,并且不需要占用本来可以用于另一个生物样本的通道。此外,我们引入了一种简单的统计方法来将来自多个iTRAQ实验的蛋白质组数据与数字响应相关联,并表明该方法比传统的基于主池的方法更有效。我们使用了四个重复的iTRAQ实验的数据来说明我们的方法,这些实验来自相同的血浆样本池中的等量,以及一个406个样本的项目,该项目旨在识别与南亚慢性营养不良儿童的营养浓度相关的血浆蛋白。
Isobaric tags for relative and absolute quantitation (iTRAQ) is a prominent mass spectrometry technology for protein identification and quantification that is capable of analyzing multiple samples in a single experiment. Frequently, iTRAQ experiments are carried out using an aliquot from a pool of all samples, or “masterpool”, in one of the channels as a reference sample standard to estimate protein relative abundances in the biological samples and to combine abundance estimates from multiple experiments. In this manuscript, we show that using a masterpool is counterproductive. We obtain more precise estimates of protein relative abundance by using the available biological data instead of the masterpool and do not need to occupy a channel that could otherwise be used for another biological sample. In addition, we introduce a simple statistical method to associate proteomic data from multiple iTRAQ experiments with a numeric response and show that this approach is more powerful than the conventionally employed masterpool-based approach. We illustrate our methods using data from four replicate iTRAQ experiments on aliquots of the same pool of plasma samples and from a 406-sample project designed to identify plasma proteins that covary with nutrient concentrations in chronically undernourished children from South Asia.
iquantitator:使用itraq进行蛋白质表达推断的工具。
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