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Review
. 2020 Nov;19(11):1739-1748.
doi: 10.1074/mcp.R120.002234. Epub 2020 Aug 26.

Single-cell Proteomics: Progress and Prospects

Ryan T Kelly  1
Affiliations
Review

Single-cell Proteomics: Progress and Prospects

Ryan T Kelly. Mol Cell Proteomics. 2020 Nov.

Abstract

MS-based proteome profiling has become increasingly comprehensive and quantitative, yet a persistent shortcoming has been the relatively large samples required to achieve an in-depth measurement. Such bulk samples, typically comprising thousands of cells or more, provide a population average and obscure important cellular heterogeneity. Single-cell proteomics capabilities have the potential to transform biomedical research and enable understanding of biological systems with a new level of granularity. Recent advances in sample processing, separations and MS instrumentation now make it possible to quantify >1000 proteins from individual mammalian cells, a level of coverage that required an input of thousands of cells just a few years ago. This review discusses important factors and parameters that should be optimized across the workflow for single-cell and other low-input measurements. It also highlights recent developments that have advanced the field and opportunities for further development.

Keywords: Cell biology; Cell sorting; FAIMS; Label-free quantification; Mass Spectrometry; Tandem Mass Spectrometry; Tissues*; nanoLC; nanoPOTS; single-cell; ultrasensitive.

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Conflict of interest statement

Conflict of interest—Author declares no competing interests.

Figures

None
Graphical abstract
Fig. 1.
Fig. 1.
Strategies for sample preparation of low input samples based on (A) iPAD, (B) OAD, (C) nanoPOTS and (d) SCoPE-MS. Adapted with permission from References , , , and , respectively.
Fig. 2.
Fig. 2.
Separation methods for single cells and other trace samples. A, Narrow-bore packed LC columns. B, Porous layer open tubular columns. C, Capillary electrophoresis with electrokinetically driven sheath flow interface. D, Nanowell-mediated 2D LC. Adapted with permission from References , , and , respectively.
See this image and copyright information in PMC

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