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Updated: Sep 14, 2025

Particle Templated Emulsification enables Microfluidic-Free Droplet Assays
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Hydrogel particle-based protein display enabled by particle-templated emulsification.

Han Wu1, Jiayao Fang1, Jiao Chen1

  • 1Institute of Chemical Biology, Shenzhen Bay Laboratory Shenzhen China wuhan@szbl.ac.cn bozheng@szbl.ac.cn.

RSC Advances
|July 24, 2025
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Summary

This study introduces a novel hydrogel particle-based protein display system using particle-templated emulsification for efficient protein discovery and engineering. This method overcomes limitations of traditional techniques, enabling rapid, compartmentalized screening with stable genotype-phenotype linkage.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Chemical Engineering

Background:

  • Conventional protein display methods, including in vivo and in vitro systems, face significant challenges.
  • In vivo methods struggle with inefficient gene transformation and complex cell dynamics.
  • In vitro methods are limited by affinity-based selection and expression bias due to homogeneous conditions.

Purpose of the Study:

  • To develop a novel hydrogel particle-based protein display technology.
  • To overcome the limitations of existing protein display and screening methods.
  • To enable high-throughput screening and protein engineering with improved efficiency and scalability.

Main Methods:

  • Particle-templated emulsification using functionalized polyacrylamide hydrogel particles as microreactors.
  • Immobilization of DNA primers for genotype and Ni-NTA groups for histidine-tagged protein phenotypes.
  • Cell-free protein expression within isolated droplets for compartmentalized screening.
  • Digital PCR for DNA amplification and stable genotype-phenotype linkage establishment.

Main Results:

  • Achieved rapid encapsulation of single hydrogel particles with DNA templates into droplets within 30 seconds.
  • Demonstrated higher single-particle encapsulation and one-to-one particle-DNA pairing efficiency compared to conventional droplet microfluidics.
  • Established a stable genotype-phenotype linkage by immobilizing amplified DNA and expressed protein on the same particle.
  • Enabled compartmentalized functional screening without cell handling, reducing reagent consumption and DNA library loss.

Conclusions:

  • The developed hydrogel particle-based protein display method is fast, scalable, and user-friendly.
  • This approach overcomes key limitations of existing in vivo and in vitro display technologies.
  • Offers significant potential for advancing directed evolution and protein engineering applications.