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

ABSTRACT: 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.

SUBMITTER: Herbrich SM

PROVIDER: S-EPMC4223774 | biostudies-literature | 2013 Feb

REPOSITORIES: biostudies-literature

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

Herbrich Shelley M SM Cole Robert N RN West Keith P KP Schulze Kerry K Yager James D JD Groopman John D JD Christian Parul P Wu Lee L O'Meally Robert N RN May Damon H DH McIntosh Martin W MW Ruczinski Ingo I

Journal of proteome research 20130116 2

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 ex ...[more]

PMID: 23270375

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Project description:Establishing the invariance property of an instrument (e.g., a questionnaire or test) is a key step for establishing its measurement validity. Measurement invariance is typically assessed by differential item functioning (DIF) analysis, i.e., detecting DIF items whose response distribution depends not only on the latent trait measured by the instrument but also on the group membership. DIF analysis is confounded by the group difference in the latent trait distributions. Many DIF analyses require knowing several anchor items that are DIF-free in order to draw inferences on whether each of the rest is a DIF item, where the anchor items are used to identify the latent trait distributions. When no prior information on anchor items is available, or some anchor items are misspecified, item purification methods and regularized estimation methods can be used. The former iteratively purifies the anchor set by a stepwise model selection procedure, and the latter selects the DIF-free items by a LASSO-type regularization approach. Unfortunately, unlike the methods based on a correctly specified anchor set, these methods are not guaranteed to provide valid statistical inference (e.g., confidence intervals and p-values). In this paper, we propose a new method for DIF analysis under a multiple indicators and multiple causes (MIMIC) model for DIF. This method adopts a minimal [Formula: see text] norm condition for identifying the latent trait distributions. Without requiring prior knowledge about an anchor set, it can accurately estimate the DIF effects of individual items and further draw valid statistical inferences for quantifying the uncertainty. Specifically, the inference results allow us to control the type-I error for DIF detection, which may not be possible with item purification and regularized estimation methods. We conduct simulation studies to evaluate the performance of the proposed method and compare it with the anchor-set-based likelihood ratio test approach and the LASSO approach. The proposed method is applied to analysing the three personality scales of the Eysenck personality questionnaire-revised (EPQ-R).

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

Publications

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

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