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Related Experiment Videos

A miniaturized arrayed assay format for detecting small molecule-protein interactions in cells

A J You1, R J Jackman, G M Whitesides

  • 1Howard Hughes Medical Institute, Department of Chemistry, Harvard University, Cambridge, MA 02138, USA.

Chemistry & Biology
|March 7, 1998
PubMed
Summary

This study introduces a miniaturized cell-based screening method for identifying protein-altering small molecules synthesized via split-pool synthesis. This technique enables rapid screening of millions of compounds in yeast and mammalian cells.

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Area of Science:

  • Biochemistry and Molecular Biology
  • Chemical Biology
  • Microfluidics and Lab-on-a-Chip Technology

Background:

  • Studying protein function involves gene mutation or small molecule binding.
  • Split-pool synthesis generates millions of unique ligands on beads, posing screening challenges.
  • Existing methods are unsuitable for large libraries of immobilized compounds, necessitating miniaturization.

Purpose of the Study:

  • To develop a miniaturized, cell-based screening technique for ligands synthesized by split-pool synthesis.
  • To enable efficient screening of large compound libraries for modulators of protein function.
  • To adapt high-throughput screening methods for covalently immobilized compounds.

Main Methods:

  • Utilized photolithography and polymer molding to create plastic devices with spatially defined nanodroplets (50-150 nL).

Related Experiment Videos

  • Developed a miniaturized, cell-based assay format compatible with yeast and mammalian cells.
  • Demonstrated detection of biologically active small molecules after photorelease from synthesis beads.

    • Main Results:

    • Successfully arrayed approximately 6,500 assays within a standard 10 cm petri dish.
    • Showcased the ability to screen ligands prepared by split-pool synthesis using microtechnology.
    • Validated the detection of small molecule effects on cellular function in the miniaturized format.

    Conclusions:

    • The described miniaturized format enables uniform nanodroplet arraying on plastic devices.
    • The spatially arrayed format ensures controlled ligand concentrations and facilitates assay automation.
    • This screening method efficiently evaluates large numbers of split-pool synthesized ligands in both yeast and mammalian cells.