Kreimer Simion, Binek Aleksandra, Chazarin Blandine, Cho Jae Hyung, Haghani Ali, Hutton Alexandre, Marbán Eduardo, Mastali Mitra, Meyer Jesse G, Mesquita Thassio, Song Yang, Van Eyk Jennifer, Parker Sarah
bioRxiv. 2023 May 31:2023.01.06.522908. doi: 10.1101/2023.01.06.522908.
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 minutes of total run time per cell with peptides quantified over 11.5 minutes 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 1,000 proteins in individual cardiomyocytes and heterogenous populations of single cells from aorta.
在复杂器官衍生的细胞混合物中识别稀有细胞类型并进行蛋白质组学表征，最好通过无标记定量质谱法来完成。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，我们展示了并行化纳流双阱单柱液相色谱（nanoDTSC），每个细胞的总运行时间为15分钟，使用标准商业组件在11.5分钟内对肽进行定量，从而提供了一种便捷高效的液相色谱解决方案，每天可分析96个单细胞。在这种通量下，nanoDTSC对单个心肌细胞和来自主动脉的单细胞异质群体中的1000多种蛋白质进行了定量。
