Division of Chemistry I, Department of Medical Biochemistry & Biophysics, Karolinska Institutet, Stockholm, Sweden.
Methods Mol Biol. 2022;2420:63-72. doi: 10.1007/978-1-0716-1936-0_6.
Single-cell proteomics is a novel application area of bioanalysis aiming to characterize proteomes of isolated single cells, which in contrast to bulk cell analysis has the potential to reveal a more detailed heterogeneity of cell populations. Although several antibody-based targeted approaches have been readily available for single-cell analysis, so far only the mass spectrometry methodology can offer unbiased proteome profiling. While this strategy has only recently emerged, it has already demonstrated unparalleled analytical power quantifying >1000 proteins in single cells. Several applications of a general isobaric labeling scheme for multiplexed sample preparation and data acquisition have been outlined using various cell types and instrumentation. This chapter provides a typical example of mass spectrometry-based single-cell proteomics workflow with details about the critical steps of analysis and alternative methods useful for optimization purposes.
单细胞蛋白质组学是生物分析的一个新的应用领域，旨在描述分离的单细胞的蛋白质组，与批量细胞分析相比，它有可能揭示细胞群体更详细的异质性。尽管已经有几种基于抗体的靶向方法可用于单细胞分析，但到目前为止，只有质谱法才能提供无偏的蛋白质组分析。虽然这种策略最近才出现，但它已经证明了无与伦比的分析能力，可以在单个细胞中定量超过 1000 种蛋白质。已经使用各种细胞类型和仪器概述了通用等压标记方案在多路样品制备和数据采集方面的一些应用。本章提供了一个基于质谱的单细胞蛋白质组学工作流程的典型示例，其中详细介绍了分析的关键步骤和用于优化目的的替代方法。
