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Application of Droplet-Array Sandwiching Technology to Click Reactions for High-Throughput Screening.

Yoshinori Miyata1, Shoma Nishimura2, Sora Kawakami2

  • 1Graduate School of Science and Engineering, Ritsumeikan University, 1-1-1, Noji-higashi, Kusatsu 525-8577, Shiga, Japan.

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|November 27, 2025
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Summary

Droplet-array sandwiching technology (DAST) combined with click chemistry offers a reagent-efficient, cross-contamination-resistant platform for high-throughput screening (HTS). This approach is well-suited for drug discovery assays, requiring minimal instrumentation.

Keywords:
click chemistrydroplet microarraydroplet-array sandwiching technologyhigh-throughput screeningmicrofluidicswettability patterning

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Area of Science:

  • Biochemistry
  • Chemical Biology
  • Drug Discovery

Background:

  • High-throughput screening (HTS) is critical in drug discovery, demanding efficient, economical, and contamination-resistant platforms.
  • Click chemistry offers biocompatibility, selectivity, and fluorescent readouts suitable for HTS.
  • Droplet-array sandwiching technology (DAST) utilizes vertically opposed droplet microarrays (DMAs) for reagent mixing and solute transport.

Purpose of the Study:

  • To integrate click chemistry with DAST for HTS applications.
  • To evaluate the feasibility of DAST as a click chemistry-based HTS platform.
  • To develop a reagent-efficient and cross-contamination-resistant HTS foundation.

Main Methods:

  • Immobilization of dibenzocyclooctyne (DBCO) on wettability-patterned (WP) substrates.
  • Verification of DBCO-azide click reaction using fluorescent azide dye.
  • Application of acoustic mixing for concentration gradient generation within DMAs.
  • Combining DAST gradient formation with click reaction fluorescence readout.

Main Results:

  • The DBCO-azide click reaction demonstrated concentration-dependent fluorescence saturation.
  • Acoustic mixing enabled independent droplet manipulation and concentration gradient formation.
  • DAST successfully reproduced concentration-dependent fluorescence, validating the approach.
  • The integrated system showed reagent efficiency and cross-contamination resistance.

Conclusions:

  • DAST is a viable HTS platform for click chemistry assays.
  • The technology offers reagent economy and resistance to cross-contamination.
  • DAST provides a foundation for low-instrument-dependent HTS assays in drug discovery.