4.5 Review

Automation of single-cell proteomic sample preparation

Journal

PROTEOMICS
Volume 21, Issue 23-24, Pages -

Publisher

WILEY
DOI: 10.1002/pmic.202100198

Keywords

automation; microfluidics; miniaturisation; robotics; sample preparation; single-cell proteomics

Funding

  1. Research Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic [ITMS2014+: 313011V446]
  2. Slovak Research and Development Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic [APVV-19-0476]
  3. Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) [404521405, SFB 1389]

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Molecular heterogeneity in biological samples is crucial for understanding pathologies and therapy resistance. Single-cell analysis using mass spectrometry can overcome issues of macroproteomics and provide more accurate insights into proteomes.
Molecular heterogeneity exists at different spatial scales in biological samples and is an important parameter in the development of pathologies and resistances to therapies. When aiming to reach molecular heterogeneity of cells at extremely low spatial scales, single-cell analysis can be the ultimate choice. Proteomics performed in bulk population of cells (macroproteomics) is prone to mask molecular heterogeneity. Mass spectrometry-based single cell proteomics (SCP-MS) is the right solution to overcome this issue. Three main problems can be identified using SCP-MS: (i) analytical loss during sample preparation, (ii) inefficient microinjection/delivery of proteins/peptides from samples to MS and (iii) low analytical throughput. Technologies for automation of SCP have recently gained attention to improve methods accuracy, sensitivity, throughput and in-depth and low-biased proteome analysis. In this minireview, we therefore overview the state-of-the-art of automation of SCP-MS sample preparation approaches.

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