In-Depth Mass Spectrometry-Based Single-Cell and Nanoscale Proteomics

Methods Mol Biol. 2021:2185:159-179. doi: 10.1007/978-1-0716-0810-4_10.

Abstract

Leukemic stem cells are highly dynamic and heterogeneous. Analysis of leukemic stem cells at the single-cell level should provide a wealth of insights that would not be possible using bulk measurements. Mass spectrometry (MS)-based proteomic workflows can quantify hundreds or thousands of proteins from a biological sample and has proven invaluable for biomedical research, but samples comprising large numbers of cells are typically required due to limited sensitivity. Recent developments in sample processing, chromatographic separations, and MS instrumentation are now extending in-depth proteome profiling to single mammalian cells. Here, we describe specific techniques that increase the sensitivity of single-cell proteomics by orders of magnitude, enabling the promise of single-cell proteomics to become a reality. We anticipate such techniques can significantly advance the understanding of leukemic stem cells.

Keywords: Proteome mapping; Single-cell analysis; Single-cell proteomics; Small cell populations; Small sample; Ultrasensitive; nanoLC; nanoPOTS.

MeSH terms

  • Humans
  • Leukemia / metabolism*
  • Mass Spectrometry*
  • Neoplastic Stem Cells / metabolism*
  • Proteomics*
  • Single-Cell Analysis*