Robust surfactant-assisted one-pot sample preparation for label-free single-cell and nanoscale proteomics

Book · Sat Jun 22 04:00:00 EDT 2024

DOI:https://doi.org/10.1007/978-1-0716-3934-4_8· OSTI ID:2477155

Xu, Zhangyang ^[1]; Zou, Rongge ^[1]; Horn, Nina C. ^[1]; Kitata, Reta Birhanu ^[1]; Shi, Tujin ^[1]

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1. BATTELLE (PACIFIC NW LAB)

With advanced mass spectrometry (MS)-based proteomics, genome-scale proteome coverage can be achieved from bulk cells. However, such bulk measurement obscures cell to cell heterogeneity, precluding proteome profiling of single cells and small numbers of cells of interest. To address this issue, in recent 5 years there are a surge of small sample preparation methods developed for robust effective collection and processing of single cells and small numbers of cells for in-depth MS-based proteome profiling. Based on their broad accessibility, they can be categorized into two types: specific device- and standard PCR tube- or multi-well plate-based methods. Herein we describe the detailed protocol of our recently developed, easily adoptable, Surfactant-assisted One-Pot (SOP) sample preparation coupled with MS method termed SOP-MS for label-free single-cell and nanoscale proteomics. SOP-MS capitalizes on the combination of a MS-compatible surfactant, DDM (n-Dodecyl-ß-D-maltoside), and standard low-bind PCR tube or multi-well plate for ‘all-in-one’ one-pot sample preparation without sample transfer. With its robust and convenient features, SOP-MS can be readily implemented in any MS laboratory for single-cell and nanoscale proteomics. With further improvements in MS detection sensitivity and sample throughput, we believe that SOP-MS could open an avenue for single-cell proteomics with broad applicability in the biological and biomedical research.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 2477155
• Report Number(s): PNNL-SA-191349
• Country of Publication: United States
• Language: English

Related Subjects

Single-cell proteomics
nanoscale proteomics
SOP-MS
surfactant
Mass Spectometry