转录组
生物
蛋白质组
单细胞分析
蛋白质组学
系统生物学
细胞
细胞生物学
计算生物学
质谱法
生物信息学
生物化学
化学
色谱法
基因
基因表达
作者
Andreas-David Brunner,Marvin Thielert,Catherine G. Vasilopoulou,Constantin Ammar,Fabian Coscia,Andreas Mund,Ole Hoerning,Nicolai Bache,Amalia Apalategui,Markus Lübeck,Sabrina Richter,David Fischer,Oliver Raether,Melvin A. Park,Florian Meier,Fabian J. Theis,Matthias Mann
标识
DOI:10.15252/msb.202110798
摘要
Single-cell technologies are revolutionizing biology but are today mainly limited to imaging and deep sequencing. However, proteins are the main drivers of cellular function and in-depth characterization of individual cells by mass spectrometry (MS)-based proteomics would thus be highly valuable and complementary. Here, we develop a robust workflow combining miniaturized sample preparation, very low flow-rate chromatography, and a novel trapped ion mobility mass spectrometer, resulting in a more than 10-fold improved sensitivity. We precisely and robustly quantify proteomes and their changes in single, FACS-isolated cells. Arresting cells at defined stages of the cell cycle by drug treatment retrieves expected key regulators. Furthermore, it highlights potential novel ones and allows cell phase prediction. Comparing the variability in more than 430 single-cell proteomes to transcriptome data revealed a stable-core proteome despite perturbation, while the transcriptome appears stochastic. Our technology can readily be applied to ultra-high sensitivity analyses of tissue material, posttranslational modifications, and small molecule studies from small cell counts to gain unprecedented insights into cellular heterogeneity in health and disease.
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