https://scholars.lib.ntu.edu.tw/handle/123456789/638331
標題: | Surfactant-assisted one-pot sample preparation for label-free single-cell proteomics | 作者: | Tsai, Chia-Feng Zhang, Pengfei Scholten, David Martin, Kendall Wang, Yi-Ting Zhao, Rui Chrisler, William B Patel, Dhwani B Dou, Maowei Jia, Yuzhi Reduzzi, Carolina Liu, Xia Moore, Ronald J Burnum-Johnson, Kristin E MIAO-HSIA LIN Hsu, Chuan-Chih Jacobs, Jon M Kagan, Jacob Srivastava, Sudhir Rodland, Karin D Steven Wiley, H Qian, Wei-Jun Smith, Richard D Zhu, Ying Cristofanilli, Massimo Liu, Tao Liu, Huiping Shi, Tujin |
公開日期: | 1-三月-2021 | 卷: | 4 | 期: | 1 | 來源出版物: | Communications biology | 摘要: | Large numbers of cells are generally required for quantitative global proteome profiling due to surface adsorption losses associated with sample processing. Such bulk measurement obscures important cell-to-cell variability (cell heterogeneity) and makes proteomic profiling impossible for rare cell populations (e.g., circulating tumor cells (CTCs)). Here we report a surfactant-assisted one-pot sample preparation coupled with mass spectrometry (MS) method termed SOP-MS for label-free global single-cell proteomics. SOP-MS capitalizes on the combination of a MS-compatible nonionic surfactant, n-Dodecyl-β-D-maltoside, and hydrophobic surface-based low-bind tubes or multi-well plates for 'all-in-one' one-pot sample preparation. This 'all-in-one' method including elimination of all sample transfer steps maximally reduces surface adsorption losses for effective processing of single cells, thus improving detection sensitivity for single-cell proteomics. This method allows convenient label-free quantification of hundreds of proteins from single human cells and ~1200 proteins from small tissue sections (close to ~20 cells). When applied to a patient CTC-derived xenograft (PCDX) model at the single-cell resolution, SOP-MS can reveal distinct protein signatures between primary tumor cells and early metastatic lung cells, which are related to the selection pressure of anti-tumor immunity during breast cancer metastasis. The approach paves the way for routine, precise, quantitative single-cell proteomics. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/638331 | ISSN: | 2399-3642 | DOI: | 10.1038/s42003-021-01797-9 |
顯示於: | 微生物學科所 |
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