通过快速循环诊断-PASEF方法的保留时间下方采样来增强识别和量化
Christopher R Below1, Oliver M Bernhardt1, Stephanie Kaspar-Schönefeld2
1Biognosys AG, Schlieren, Switzerland.
Molecular & cellular proteomics : MCP
|December 11, 2025
概括
在timsTOF HT上优化对角-PASEF与保留时间总和 (RTsum) 显著提高了蛋白质组量化. 与标准方法相比,这种新的方法可以改善信号噪声比率,并增加和蛋白质的识别.
科学领域:
- 质谱测量质量谱测量
- 蛋白质组学是指蛋白质组学
- 分析化学是一种分析化学.
背景情况:
- 数据独立采集 (DIA) 质谱对于全面的蛋白质组量化至关重要.
- 诊断-PASEF是timsTOF平台上的一个先进的获取方法,用于改进前体离子分析.
- 对于常规应用,需要进一步优化对角-PASEF工作流程.
研究的目的:
- 系统地优化对角-PASEF在17分钟梯度上的时间TOF HT上的获取.
- 通过将对角-PASEF与保留时间总和 (RTsum) 结合起来,开发和验证一种增强的分析策略.
- 为了比较优化的对角-PASEF策略与标准的对角-PASEF方法的性能.
主要方法:
- 在 timsTOF HT仪器上对对角-PASEF采集参数的系统优化.
- 将保留时间总和 (RTsum) 与对角-PASEF数据采集集成.
- 使用 Spectronaut 软件进行数据分析的性能评估.
- 用细胞系样本的不同输入量进行定量实验.
主要成果:
- "光谱探测器"完全支持对角-PASEF方法,用户干预最小.
- RTsum与对角PASEF的结合提高了信号噪声比和峰值形状.
- 这种综合策略显著增加了和蛋白质识别的数量,而不是标准的PASEF.
- 用RTsum进行的Diagonal-PASEF在受控实验中表现出卓越的定量精度和识别率.
结论:
- 与RTsum相结合的Diagonal-PASEF代表了深度蛋白质组定量化的强大策略.
- 这种优化的工作流提供了对现有的DIA方法的有竞争力的替代方案,提高了识别深度和定量准确性.
- RTsum有效地利用了对角-PASEF的高采样率,提高了其在蛋白质组分析中的实用性.
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