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A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
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A High-Throughput Yeast Halo Assay for Bioactive Compounds.

Walter Bray1, R Scott Lokey1

  • 1Department of Chemistry and Biochemistry, University of California Santa Cruz, Santa Cruz, California 95064.

Cold Spring Harbor Protocols
|September 3, 2016
PubMed
Summary

A high-throughput halo assay was developed for drug screening. This method automates the traditional yeast-based assay, enabling rapid quantification of drug potency and biological activity for thousands of compounds daily.

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Area of Science:

  • Microbiology
  • Drug Discovery
  • Assay Development

Background:

  • The traditional halo assay is a long-standing method for assessing antifungal and antibiotic activity.
  • This assay relies on measuring clear zones around drug-impregnated disks on yeast-seeded agar plates.
  • However, the conventional method is low-throughput and material-intensive.

Purpose of the Study:

  • To develop a high-throughput version of the halo assay for efficient drug screening.
  • To enable automated quantification and data archiving of biological activity.
  • To adapt the established halo assay for large-scale compound evaluation.

Main Methods:

  • A 384-pin array was utilized to dispense small volumes (∼200 nL) of compound stock solutions.
  • Stock solutions were applied to single-well plates pre-seeded with yeast.
  • Resulting halos were quantified using a plate reader in absorbance mode.

Main Results:

  • The high-throughput halo assay successfully quantified yeast growth inhibition zones.
  • The assay demonstrated automation capabilities for screening thousands of compounds per day.
  • Data generated were archived in flat files, compatible with compound databases.

Conclusions:

  • The developed high-throughput halo assay offers a scalable and efficient alternative to traditional methods.
  • This automated assay retains the visual readout convenience while significantly increasing screening capacity.
  • The protocol facilitates rapid assessment of cytotoxic or cytostatic drug potency against yeast.