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Phage-Based Profiling of Rare Single Cells Using Nanoparticle-Directed Capture.

Yuan Ma1,2, Kangfu Chen1, Fan Xia1

  • 1Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, Ontario M5S 3M2, Canada.

ACS Nano
|November 23, 2021
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Summary

This study introduces a microfluidic chip for analyzing rare circulating tumor cells (CTCs). The platform enables high-efficiency capture and single-cell protein analysis, revealing microenvironment-influenced Frizzled2 expression changes.

Keywords:
barcoded phage-displayed antibody screeningcirculating tumor cellsmicrofluidicsphage-based profilingprotein expressionsingle cell

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Area of Science:

  • Biotechnology
  • Proteomics
  • Microfluidics

Background:

  • Quantitative proteomic analysis at the single-cell level is crucial for rare cell research, including circulating tumor cell (CTC) profiling.
  • Existing platforms often lack the sensitivity and versatility required for detailed analysis of rare cell populations.

Purpose of the Study:

  • To develop and validate an integrated microfluidic platform for high-efficiency capture and in situ analysis of single circulating tumor cells.
  • To investigate dynamic changes in cell-surface protein expression in response to adherence and microenvironment cues at single-cell resolution.

Main Methods:

  • An integrated microfluidic chip utilizing magnetic nanoparticles for efficient single tumor cell capture.
  • On-chip incubation and in situ analysis of cell-surface protein expression.
  • Phage-based barcoding coupled with next-generation sequencing for multiplexed surface marker monitoring.

Main Results:

  • Demonstrated high-efficiency capture of single tumor cells using the microfluidic chip.
  • Successfully monitored dynamic changes in multiple cell-surface markers upon CTC adherence.
  • Observed fluctuations in Frizzled2 (FZD2) expression correlating with the single-cell microenvironment.

Conclusions:

  • The developed microfluidic platform offers a powerful tool for in-depth, single-cell resolution screening of multiple surface antigens in rare cells.
  • This technology provides valuable insights into biological heterogeneity and disease mechanisms by analyzing dynamic protein expression.
  • Potential applications include advanced diagnostics and therapeutic target identification for various human diseases.