Leibniz-Institut für Analytische Wissenschaften - ISAS - e.V., Dortmund, Germany.
Proteomics. 2011 Mar;11(6):1125-34. doi: 10.1002/pmic.201000711. Epub 2011 Feb 16.
Reporter ion-based methods are among the major techniques to quantify peptides and proteins. Two main labels, tandem mass tag (TMT) and iTRAQ, are widely used by the proteomics community. They are, however, often applied as out-of-the-box methods, without thorough quality control. Thus, due to undiscovered limitations of the technique, irrelevant results might be trusted. To address this issue, we here propose a step-by-step quality control of the iTRAQ workflow. From sample preparation to final ratio calculation we provide metrics and techniques assessing the actual effectiveness of iTRAQ quantification as well as a novel method for more reliable protein ratio estimation.
基于离子的方法是定量肽和蛋白质的主要技术之一。串联质量标签（TMT）和 iTRAQ 是蛋白质组学领域广泛使用的两种主要标签。然而，它们通常作为开箱即用的方法应用，而没有进行彻底的质量控制。因此，由于技术的未发现的局限性，可能会信任不相关的结果。为了解决这个问题，我们在这里提出了 iTRAQ 工作流程的逐步质量控制。从样品制备到最终比率计算，我们提供了评估 iTRAQ 定量实际有效性的指标和技术，以及一种更可靠的蛋白质比率估计的新方法。
