Data-independent acquisition modes isolate and concurrently fragment populations of different precursors by cycling through segments of a predefined precursor m/z range. Although these selection windows collectively cover the entire m/z range, overall, only a few per cent of all incoming ions are isolated for mass analysis. Here, we make use of the correlation of molecular weight and ion mobility in a trapped ion mobility device (timsTOF Pro) to devise a scan mode that samples up to 100% of the peptide precursor ion current in m/z and mobility windows. We extend an established targeted data extraction workflow by inclusion of the ion mobility dimension for both signal extraction and scoring and thereby increase the specificity for precursor identification. Data acquired from whole proteome digests and mixed organism samples demonstrate deep proteome coverage and a high degree of reproducibility as well as quantitative accuracy, even from 10 ng sample amounts.

与数据无关的采集模式通过循环通过预定义的前体m / z范围的片段来隔离不同前体的种群，并同时使其片段化。尽管这些选择窗口共同覆盖了整个m / z范围，但总的来说，仅隔离了所有进入离子中的百分之几进行质量分析。在这里，我们利用捕获的离子淌度设备（timsTOF Pro）中分子量和离子淌度的相关性来设计一种扫描模式，以m / z为单位对高达100％的肽前体离子电流进行采样和移动窗口。我们通过包含离子迁移率维度来扩展既定的目标数据提取工作流程，以进行信号提取和评分，从而提高了前体识别的特异性。从整个蛋白质组消化物和混合的生物样品中获取的数据表明，即使从10 ng样品量中提取，蛋白质组覆盖面也很广，具有很高的重现性和定量准确性。