ChELSI Institute, Department of Chemical and Biological Engineering, University of Sheffield, Sheffield, UK.
Proteomics. 2011 Jun;11(11):2341-6. doi: 10.1002/pmic.201000752. Epub 2011 May 4.
Application of iTRAQ-based workflows for protein profiling has become widespread. Concomitantly, the idiosyncratic limitations of iTRAQ, such as its tendency to underestimate quantifications, have been studied and recognised. This report shows that the influence of ratio compression and limiting transmission in iTRAQ MS/MS in high-complexity mixtures (iTRAQ-labelled lysates) can be partly alleviated using high-resolution sample fractionation. Here, we also investigate in greater detail the dependency of iTRAQ quantification on the dynamics of online chromatography in low-complexity mixtures (iTRAQ-labelled standards). These findings will allow more efficient strategies to be designed for iTRAQ proteomics, alleviating iTRAQ underestimation and thus facilitating the detection of subtle abundance changes.
基于 iTRAQ 的蛋白质谱分析技术已经得到广泛应用。与此同时，iTRAQ 技术也存在一些固有局限性，例如定量时容易低估，这些局限性已得到研究和认可。本报告显示，使用高分辨率样品分离可部分缓解高复杂度混合物（iTRAQ 标记的裂解物）中 iTRAQ MS/MS 中比率压缩和限制传输的影响。在这里，我们还更详细地研究了 iTRAQ 定量对低复杂度混合物（iTRAQ 标记标准品）中在线色谱动力学的依赖性。这些发现将有助于设计更有效的 iTRAQ 蛋白质组学策略，减轻 iTRAQ 低估，从而更易于检测微妙的丰度变化。
