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