Surfactant-assisted one-pot sample preparation for label-free single-cell proteomics.


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.

  • Burnum-Johnson KE
  • Chrisler WB
  • Cristofanilli M
  • Dou M
  • Hsu CC
  • Jacobs JM
  • Jia Y
  • Kagan J
  • Lin MH
  • Liu H
  • Liu T
  • Liu X
  • Martin K
  • Moore RJ
  • Patel DB
  • Qian WJ
  • Reduzzi C
  • Rodland KD
  • Scholten D
  • Shi T
  • Smith RD
  • Srivastava S
  • Steven Wiley H
  • Tsai CF
  • Wang YT
  • Zhang P
  • Zhao R
  • Zhu Y
PubMed ID
Appears In
Commun Biol, 2021, 4 (1)