丹麦哥本哈根大学Jesper V. Olsen研究小组开发出新型蛋白组质谱技术。2024年2月1日，国际知名学术期刊《自然—生物技术》在线发表了这一成果。

研究人员介绍了窄窗数据独立采集（nDIA）策略，包括高分辨率MS1扫描和使用2-Th隔离窗的约200Hz并行串联质谱（MS/MS）扫描，消除了数据独立采集和数据独立采集方法之间的差异。这是通过将四极杆Orbitrap质谱仪与非对称轨迹无损（Astral）分析仪配对实现的，后者提供大于200Hz的MS/MS扫描速度、高分辨率和灵敏度以及低ppm质量精度。

采用nDIA策略，每天可分析>100个完整的酵母蛋白质组，或每天分析48个人类蛋白质组，半小时内可分析约10000个人类蛋白质组，5分钟内可分析约7000个蛋白质，与目前最先进的质谱（MS）相比，覆盖率提高了3倍。对离线分馏样品进行多拍采集，可在约3小时内全面覆盖人类蛋白质组。在三物种蛋白质组混合物中，单次半小时运行可量化14000多个蛋白质组，显示了高定量精度和准确性。

研究人员表示，基于MS的蛋白质组学旨在以快速、可重复的方式描述综合蛋白质组的特征。

附：英文原文

Title: Ultra-fast label-free quantification and comprehensive proteome coverage with narrow-window data-independent acquisition

Author: Guzman, Ulises H., Martinez-Val, Ana, Ye, Zilu, Damoc, Eugen, Arrey, Tabiwang N., Pashkova, Anna, Renuse, Santosh, Denisov, Eduard, Petzoldt, Johannes, Peterson, Amelia C., Harking, Florian, stergaard, Ole, Rydbirk, Rasmus, Aznar, Susana, Stewart, Hamish, Xuan, Yue, Hermanson, Daniel, Horning, Stevan, Hock, Christian, Makarov, Alexander, Zabrouskov, Vlad, Olsen, Jesper V.

Issue&Volume: 2024-02-01

Abstract: Mass spectrometry (MS)-based proteomics aims to characterize comprehensive proteomes in a fast and reproducible manner. Here we present the narrow-window data-independent acquisition (nDIA) strategy consisting of high-resolution MS1 scans with parallel tandem MS (MS/MS) scans of ~200Hz using 2-Th isolation windows, dissolving the differences between data-dependent and -independent methods. This is achieved by pairing a quadrupole Orbitrap mass spectrometer with the asymmetric track lossless (Astral) analyzer which provides >200-Hz MS/MS scanning speed, high resolving power and sensitivity, and low-ppm mass accuracy. The nDIA strategy enables profiling of >100 full yeast proteomes per day, or 48 human proteomes per day at the depth of ~10,000 human protein groups in half-an-hour or ~7,000 proteins in 5min, representing 3× higher coverage compared with current state-of-the-art MS. Multi-shot acquisition of offline fractionated samples provides comprehensive coverage of human proteomes in ~3h. High quantitative precision and accuracy are demonstrated in a three-species proteome mixture, quantifying 14,000+ protein groups in a single half-an-hour run.

DOI: 10.1038/s41587-023-02099-7

Source: https://www.nature.com/articles/s41587-023-02099-7