蛋白质组
赫拉
单细胞分析
计算生物学
工作流程
细胞
鸟枪蛋白质组学
蛋白质组学
鉴定(生物学)
化学
生物
细胞生物学
生物信息学
计算机科学
生物化学
基因
植物
数据库
作者
Yu Sheng Wang,Zhi-Ying Guan,Shao-Wen Shi,Yunxia Jiang,Jie Zhang,Yi Yang,Qiong Wu,Jie Wu,Jianbo Chen,Wei-Xin Ying,Qinqin Xu,Qian-Xi Fan,Huifeng Wang,Lin Zhou,Sheng Wang,Fang Jin,Jian-Zhang Pan,Qun Fang
标识
DOI:10.1038/s41467-024-45659-4
摘要
The shotgun proteomic analysis is currently the most promising single-cell protein sequencing technology, however its identification level of ~1000 proteins per cell is still insufficient for practical applications. Here, we develop a pick-up single-cell proteomic analysis (PiSPA) workflow to achieve a deep identification capable of quantifying up to 3000 protein groups in a mammalian cell using the label-free quantitative method. The PiSPA workflow is specially established for single-cell samples mainly based on a nanoliter-scale microfluidic liquid handling robot, capable of achieving single-cell capture, pretreatment and injection under the pick-up operation strategy. Using this customized workflow with remarkable improvement in protein identification, 2449-3500, 2278-3257 and 1621-2904 protein groups are quantified in single A549 cells (n = 37), HeLa cells (n = 44) and U2OS cells (n = 27) under the DIA (MBR) mode, respectively. Benefiting from the flexible cell picking-up ability, we study HeLa cell migration at the single cell proteome level, demonstrating the potential in practical biological research from single-cell insight.
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