Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells Academic Article uri icon

Overview

MeSH Major

  • Intracellular Signaling Peptides and Proteins
  • Protein Array Analysis
  • Proteomics
  • Signal Transduction

abstract

  • We describe a microchip designed to quantify the levels of a dozen cytoplasmic and membrane proteins from single cells. We use the platform to assess protein-protein interactions associated with the EGF-receptor-mediated PI3K signaling pathway. Single-cell sensitivity is achieved by isolating a defined number of cells (n = 0-5) in 2 nL volume chambers, each of which is patterned with two copies of a miniature antibody array. The cells are lysed on-chip, and the levels of released proteins are assayed using the antibody arrays. We investigate three isogenic cell lines representing the cancer glioblastoma multiforme, at the basal level, under EGF stimulation, and under erlotinib inhibition plus EGF stimulation. The measured protein abundances are consistent with previous work, and single-cell analysis uniquely reveals single-cell heterogeneity, and different types and strengths of protein-protein interactions. This platform helps provide a comprehensive picture of altered signal transduction networks in tumor cells and provides insight into the effect of targeted therapies on protein signaling networks.

publication date

  • January 10, 2012

Research

keywords

  • Academic Article

Identity

Language

  • eng

PubMed Central ID

  • PMC3258586

Digital Object Identifier (DOI)

  • 10.1073/pnas.1110865109

PubMed ID

  • 22203961

Additional Document Info

start page

  • 419

end page

  • 24

volume

  • 109

number

  • 2