The analytical scale of most mass‐spectrometry‐based targeted proteomics assays is usually limited by assay performance and instrument utilization. A recently introduced method, called triggered by offset, multiplexed, accurate mass, high resolution, and absolute quantitation (TOMAHAQ), combines both peptide and sample multiplexing to simultaneously improve analytical scale and quantitative performance. In the present work, critical technical requirements and data analysis considerations for successful implementation of the TOMAHAQ technique based on the study of a total of 185 target peptides across over 200 clinical plasma samples are discussed. Importantly, it is observed that significant interference originate from the TMTzero reporter ion used for the synthetic trigger peptides. This interference is not expected because only TMT10plex reporter ions from the target peptides should be observed under typical TOMAHAQ conditions. In order to unlock the great promise of the technique for high throughput quantification, here a post‐acquisition data correction strategy to deconvolute the reporter ion superposition and recover reliable data is proposed.

中文翻译：

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增强TOMAHAQ中数据的可靠性，以进行大规模蛋白质定量。

大多数基于质谱的靶向蛋白质组学测定方法的分析规模通常受到测定方法性能和仪器利用率的限制。最近引入的一种方法称为偏移，多路复用，精确质量，高分辨率和绝对定量（TOMAHAQ）触发，结合了肽和样品多路复用功能，同时提高了分析规模和定量性能。在当前的工作中，基于对200多个临床血浆样品中总共185种目标肽的研究，讨论了成功实施TOMAHAQ技术的关键技术要求和数据分析注意事项。重要的是，观察到明显的干扰源自用于合成触发肽的TMTzero报告离子。由于在典型的TOMAHAQ条件下仅应观察到来自目标肽的TMT10plex报告离子，所以不会产生这种干扰。为了释放高通量定量技术的巨大前景，在此提出了一种后卷积数据校正策略，以对报告离子叠加进行反卷积并恢复可靠的数据。