Bioinformatics Program, Boston University, Boston, MA, USA.
Department of Biochemistry and Cell Biology, Boston University, Boston, MA, USA.
Anal Bioanal Chem. 2024 Apr;416(9):2359-2369. doi: 10.1007/s00216-024-05171-6. Epub 2024 Feb 15.
Success of mass spectrometry characterization of the proteome of single cells allows us to gain a greater understanding than afforded by transcriptomics alone but requires clear understanding of the tradeoffs between analytical throughput and precision. Recent advances in mass spectrometry acquisition techniques, including updated instrumentation and sample preparation, have improved the quality of peptide signals obtained from single cell data. However, much of the proteome remains uncharacterized, and higher throughput techniques often come at the expense of reduced sensitivity and coverage, which diminish the ability to measure proteoform heterogeneity, including splice variants and post-translational modifications, in single cell data analysis. Here, we assess the growing body of ultrasensitive single-cell approaches and their tradeoffs as researchers try to balance throughput and precision in their experiments.
单细胞蛋白质组的质谱分析的成功使我们对细胞的理解程度超过了转录组学，但这需要我们对分析通量和精度之间的权衡有清晰的认识。近年来，质谱采集技术取得了进展，包括更新的仪器和样品制备，这提高了从单细胞数据中获得的肽信号的质量。然而，大部分蛋白质组仍未被描述，更高通量的技术通常是以降低灵敏度和覆盖度为代价的，这降低了在单细胞数据分析中测量蛋白质异构体异质性（包括剪接变体和翻译后修饰）的能力。在这里，我们评估了越来越多的超灵敏单细胞方法及其权衡，因为研究人员试图在实验中平衡通量和精度。
