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A Fluorescence-Based High-Throughput Screening Assay to Identify Growth Inhibitors of the Pathogenic Fungus Aspergillus fumigatus

  • Thomas M. SmithEmail author
  • Daryl L. Richie
  • Jianshi Tao
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1439)

Abstract

Due to the advancements in modern medicine that have resulted in an increased number of immunocompromised individuals, the incidences and the associated mortality of invasive aspergillosis have continued to rise over the past three decades despite appropriate treatment. As a result, invasive aspergillosis has emerged as a leading cause of infection-related mortality in immunocompromised individuals. Utilizing the resazurin to resorufin conversion fluorescence readout to monitor cell viability, herein, we outline a high-throughput screening method amenable to profiling a large pharmaceutical library against the clinically relevant but less frequently screened fungal pathogen Aspergillus fumigatus. This enables the user to conduct high-throughput screening using a disease-relevant fungal growth assay and identify novel antifungal chemotypes as drug leads.

Key words

Aspergillus fumigatus Fungus Antifungal Resazurin High-throughput screening (HTS) 

Notes

Acknowledgements

We would like to thank the Aspergillus fumigatus inhibitor project team including Neil Ryder, Kathryn Thompson, Roger Fujimoto, Joseph Drumm, and Dominic Hoepfner for scientific input and advice during the screen. We would like to thank Heidi Faw for safety considerations regarding Aspergillus handling. We would like to thank Alejandra Raimondi for initial assay development work. We would like to thank Melissa Grippo for compound purity testing by LC-MS.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Thomas M. Smith
    • 1
    Email author
  • Daryl L. Richie
    • 2
  • Jianshi Tao
    • 3
  1. 1.Center for Proteomic ChemistryNovartis Institutes for BioMedical Research, Inc.CambridgeUSA
  2. 2.Infectious Diseases AreaNovartis Institutes for BioMedical Research, Inc.EmeryvilleUSA
  3. 3.Genomics Institute of the Novartis Research FoundationSan DiegoUSA

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