Detection of Isoforms Differing by a Single Charge Unit in Individual Cells

Angew Chem Int Ed Engl. 2016 Sep 26;55(40):12431-5. doi: 10.1002/anie.201606039. Epub 2016 Sep 6.

Abstract

To measure protein isoforms in individual mammalian cells, we report single-cell resolution isoelectric focusing (scIEF) and high-selectivity immunoprobing. Microfluidic design and photoactivatable materials establish the tunable pH gradients required by IEF and precisely control the transport and handling of each 17-pL cell lysate during analysis. The scIEF assay resolves protein isoforms with resolution down to a single-charge unit, including both endogenous cytoplasmic and nuclear proteins from individual mammalian cells.

Keywords: immunoassays; isoelectric focusing; lab-on-a-chip; proteomics; single-cell analysis.

Publication types

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

MeSH terms

  • Cell Line, Tumor
  • Fluorescent Dyes / chemistry
  • Humans
  • Hydrogen-Ion Concentration
  • Immunoassay
  • Isoelectric Focusing / methods*
  • Lab-On-A-Chip Devices
  • Protein Isoforms / analysis*
  • Single-Cell Analysis

Substances

  • Fluorescent Dyes
  • Protein Isoforms