Proteomic Analysis of Single Mammalian Cells Enabled by Microfluidic Nanodroplet Sample Preparation and Ultrasensitive NanoLC-MS

Angew Chem Int Ed Engl. 2018 Sep 17;57(38):12370-12374. doi: 10.1002/anie.201802843. Epub 2018 Jun 14.

Abstract

We report on the quantitative proteomic analysis of single mammalian cells. Fluorescence-activated cell sorting was employed to deposit cells into a newly developed nanodroplet sample processing chip, after which samples were analyzed by ultrasensitive nanoLC-MS. An average of circa 670 protein groups were confidently identified from single HeLa cells, which is a far greater level of proteome coverage for single cells than has been previously reported. We demonstrate that the single-cell proteomics platform can be used to differentiate cell types from enzyme-dissociated human lung primary cells and identify specific protein markers for epithelial and mesenchymal cells.

Keywords: cell typing; droplets; microfluidics; single-cell proteomics; ultrasensitive LC-MS.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Chromatography, High Pressure Liquid / methods
  • HeLa Cells
  • Humans
  • Lung / cytology
  • Lung / metabolism
  • Microfluidics / methods*
  • Nanotechnology / methods*
  • Principal Component Analysis
  • Proteome / analysis*
  • Proteomics / methods*
  • Tandem Mass Spectrometry / methods

Substances

  • Proteome