Abstract

Single-cell functional proteomics assays can connect genomic information to biological function through quantitative and multiplex protein measurements. Tools for single-cell proteomics have developed rapidly over the past 5 years and are providing approaches for directly elucidating phosphoprotein signaling networks in cancer cells or for capturing high-resolution snapshots of immune system function in patients with various disease conditions. We discuss advances in single-cell proteomics platforms, with an emphasis on microchip methods. These methods can provide a direct correlation of morphological, functional and molecular signatures at the single-cell level. We also provide examples of how those platforms are being applied to both fundamental biology and clinical studies, focusing on immune-system monitoring and phosphoprotein signaling networks in cancer.

Highlights

  • Single-cell functional proteomics assays can connect genomic information to biological function through quantitative and multiplex protein measurements

  • Cell-to-cell variation and single-cell functional proteomics analysis Non-genetic cellular heterogeneity is a universal feature of any cell population [1,2]

  • For intracellular signaling networks, such as those associated with growth factor signaling, correlations and anti-correlations between phosphoproteins can indicate activating and inhibitory interactions, respectively

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Summary

Introduction

Single-cell functional proteomics assays can connect genomic information to biological function through quantitative and multiplex protein measurements. Miniature antibody arrays yield spatial separation of specific protein assays; proteomeic and functional assays from same cell; single cells or defined small cell populations accessed. The most advanced microfluidic single-cell proteomics tools use surface-immobilized antibodies for separating protein detection from cell manipulation (Figure 1c,d).

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