丹麦哥本哈根大学Jesper V. Olsen等研究人员合作发现，借助Chip-Tip工作流程提升的灵敏度和可扩展性可实现深度单细胞蛋白质组学。相关论文于2025年1月16日在线发表在《自然—方法学》杂志上。

研究人员提出了一种高灵敏度的单细胞蛋白质组学（SCP）工作流程——Chip-Tip，能够在单个HeLa细胞中识别超过5000种蛋白质。该方法还能够直接检测单个细胞中的翻译后修饰，无需特定的翻译后修饰富集过程。该研究展示了每天处理多达120个无标记SCP样本的可行性。

优化的组织解离缓冲液使得药物处理后的癌细胞球体，能够有效地进行单细胞分散，从而提高了整体的SCP分析效果。在分析非定向的人源诱导多能干细胞分化时，研究人员一致性地定量了，人源诱导多能干细胞中的干细胞标志物OCT4和SOX2，并在不同的胚体细胞中定量了谱系标志物，如GATA4（内胚层）、HAND1（中胚层）和MAP2（外胚层）。

该工作流程在SCP的灵敏度和通量方面设立了标杆，具有广泛的基础生物学和生物医学应用，特别是在识别特定细胞类型标志物和治疗靶点方面。

据介绍，SCP有望通过提供单个细胞中蛋白质组的无与伦比视角，从而彻底改变生物医学领域。

附：英文原文

Title: Enhanced sensitivity and scalability with a Chip-Tip workflow enables deep single-cell proteomics

Author: Ye, Zilu, Sabatier, Pierre, van der Hoeven, Leander, Lechner, Maico Y., Phlairaharn, Teeradon, Guzman, Ulises H., Liu, Zhen, Huang, Haoran, Huang, Min, Li, Xiangjun, Hartlmayr, David, Izaguirre, Fabiana, Seth, Anjali, Joshi, Hiren J., Rodin, Sergey, Grinnemo, Karl-Henrik, Hrning, Ole B., Bekker-Jensen, Dorte B., Bache, Nicolai, Olsen, Jesper V.

Issue&Volume: 2025-01-16

Abstract: Single-cell proteomics (SCP) promises to revolutionize biomedicine by providing an unparalleled view of the proteome in individual cells. Here, we present a high-sensitivity SCP workflow named Chip-Tip, identifying >5,000 proteins in individual HeLa cells. It also facilitated direct detection of post-translational modifications in single cells, making the need for specific post-translational modification-enrichment unnecessary. Our study demonstrates the feasibility of processing up to 120 label-free SCP samples per day. An optimized tissue dissociation buffer enabled effective single-cell disaggregation of drug-treated cancer cell spheroids, refining overall SCP analysis. Analyzing nondirected human-induced pluripotent stem cell differentiation, we consistently quantified stem cell markers OCT4 and SOX2 in human-induced pluripotent stem cells and lineage markers such as GATA4 (endoderm), HAND1 (mesoderm) and MAP2 (ectoderm) in different embryoid body cells. Our workflow sets a benchmark in SCP for sensitivity and throughput, with broad applications in basic biology and biomedicine for identification of cell type-specific markers and therapeutic targets.

DOI: 10.1038/s41592-024-02558-2

Source: https://www.nature.com/articles/s41592-024-02558-2