Highly multiplexed profiling of cell surface proteins on single circulating tumor cells based on antibody and cellular barcoding
Highly multiplexed profiling of cell surface proteins on single circulating tumor cells based on antibody and cellular barcoding
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基于抗体和细胞条形码对单个循环肿瘤细胞的细胞表面蛋白进行高度多重分析
DOI：
10.1007/s00216-019-01666-9
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发表时间：
2019
影响因子：
4.3
通讯作者：
Shi Qihui
中科院分区：
文献类型：
--
作者：
Wang Chunying;Yang Liu;Wang Zhuo;He Jianjun;Shi Qihui
循环肿瘤细胞(CTCs)在血液样本中非常罕见，代表了对肿瘤的实时“液体活组织检查”。虽然已经报道了单个CTCs的遗传和转录测序，但这些方法无法提供CTCs的表型和功能信息，如表面蛋白的蛋白质水平。由于缺乏单细胞蛋白质组学方法来处理和分析高背景的非靶细胞中的稀有细胞，因此对CTCs的单细胞蛋白质组分析的研究一直很少。在这里，我们发展了一种基于抗体和细胞DNA条码策略的微芯片辅助单细胞蛋白质组学方法来分析CTCs的表面蛋白。我们将DNA编码的抗体标签和细胞指数结合在一起，同时对~ 100单个稀有细胞中的15种蛋白质进行分析，并使用高通量测序作为读数，根据细胞指数和抗体衍生的蛋白条形码生成CTC的表面蛋白质谱。利用6400孔微芯片和自动打孔机，以最小的细胞损失(~ 10%)，从浓缩的四氯化碳种群中快速提取单个四氯化碳。该技术平台集成了单个CTCs的可靠分离和蛋白质组学分析，可扩展到数百个稀有细胞中的~ 100蛋白质，并具有单细胞精度。
Circulating tumor cells (CTCs) are extraordinarily rare in blood samples and represent a real-time “liquid biopsy” of tumors. Although genetic and transcriptional sequencing of single CTCs has been reported, these methods fail to provide phenotypic and functional information of CTCs such as protein levels of surface proteins. Studies of single-cell proteomic assays of CTCs have been rare because of a lack of single-cell proteomic methods to handle and analyze rare cells in a high background of non-target cells with high sensitivity, throughput, and multiplexing capacity. Here, we develop a microchip-assisted single-cell proteomic method for profiling surface proteins of CTCs based on antibody and cellular DNA barcoding strategy. We combine DNA-encoded antibody tags and cell indexes to profile 15 proteins in ~ 100 single rare cells simultaneously, and use high-throughput sequencing as the readout to generate surface protein profiles of CTCs according to their cell indexes and antibody-derived protein barcodes. A 6400-well microchip and the automated puncher are used to rapidly retrieve single CTCs from enriched CTC population with minimal cell loss (~ 10%). This technological platform integrates reliable isolation and proteomic analysis of single CTCs and can be extendable to ~ 100 proteins in hundreds of rare cells with single-cell precision.