通过纳米流双重陷阱单柱液相色谱对器官来源的异质细胞群进行高通量单细胞蛋白质组学分析。
High-Throughput Single-Cell Proteomic Analysis of Organ-Derived Heterogeneous Cell Populations by Nanoflow Dual-Trap Single-Column Liquid Chromatography.
机构信息
Advanced Clinical Biosystems Research Institute, Smidt Heart Institute, Cedars-Sinai Medical Center, Los Angeles, California 90048, United States.
Smidt Heart Institute, Cedars-Sinai Medical Center, Los Angeles, California 90048, United States.
出版信息
Anal Chem. 2023 Jun 20;95(24):9145-9150. doi: 10.1021/acs.analchem.3c00213. Epub 2023 Jun 8.
Abstract
Identification and proteomic characterization of rare cell types within complex organ-derived cell mixtures is best accomplished by label-free quantitative mass spectrometry. High throughput is required to rapidly survey hundreds to thousands of individual cells to adequately represent rare populations. Here we present parallelized nanoflow dual-trap single-column liquid chromatography (nanoDTSC) operating at 15 min of total run time per cell with peptides quantified over 11.5 min using standard commercial components, thus offering an accessible and efficient LC solution to analyze 96 single cells per day. At this throughput, nanoDTSC quantified over 1000 proteins in individual cardiomyocytes and heterogeneous populations of single cells from the aorta.
摘要
通过无标记定量质谱分析，可最好地鉴定和蛋白质组学鉴定复杂器官来源细胞混合物中的稀有细胞类型。需要高通量才能快速检测数百到数千个单个细胞，以充分代表稀有群体。在这里，我们展示了平行纳米流双阱单柱液相色谱（nanoDTSC），每个细胞的总运行时间为 15 分钟，使用标准商业组件对肽进行 11.5 分钟的定量，从而提供了一种可访问且高效的 LC 解决方案，每天可分析 96 个单细胞。在这个通量下，nanoDTSC 可对单个心肌细胞和主动脉的单细胞异质群体中的 1000 多种蛋白质进行定量。
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