Portal de Periódicos da CAPES

Pick-up single-cell proteomic analysis for quantifying up to 3000 proteins in a Mammalian cell

2024; Nature Portfolio; Volume: 15; Issue: 1 Linguagem: Inglês

10.1038/s41467-024-45659-4

2041-1723

Yu Wang, Zhi-Ying Guan, Shao-Wen Shi, Yirong Jiang, Jie Zhang, Yi Yang, Qiong Wu, Jie Wu, Jianbo Chen, Wei-Xin Ying, Qin‐Qin Xu, Qian‐Xi Fan, Hui-Feng Wang, Li Zhou, Ling Wang, Fang Jin, Jian‐Zhang Pan, Qun Fang,

Cell Image Analysis Techniques

Abstract 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.