Related Experiment Video
Updated: May 3, 2026
High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds
Published on: August 18, 2023
High-content, high-throughput screening for the identification of cytotoxic compounds based on cell morphology and
Heather L Martin1, Matthew Adams2, Julie Higgins2
1BioScreening Technology Group, Leeds Institutes of Molecular Medicine, University of Leeds, Leeds, United Kingdom ; School of Chemistry, University of Leeds, Leeds, United Kingdom.
Abstract:
Toxicity is a major cause of failure in drug discovery and development, and whilst robust toxicological testing occurs, efficiency could be improved if compounds with cytotoxic characteristics were identified during primary compound screening. The use of high-content imaging in primary screening is becoming more widespread, and by utilising phenotypic approaches it should be possible to incorporate cytotoxicity counter-screens into primary screens. Here we present a novel phenotypic assay that can be used as a counter-screen to identify compounds with adverse cellular effects. This assay has been developed using U2OS cells, the PerkinElmer Operetta high-content/high-throughput imaging system and Columbus image analysis software. In Columbus, algorithms were devised to identify changes in nuclear morphology, cell shape and proliferation using DAPI, TOTO-3 and phosphohistone H3 staining, respectively. The algorithms were developed and tested on cells treated with doxorubicin, taxol and nocodazole. The assay was then used to screen a novel, chemical library, rich in natural product-like molecules of over 300 compounds, 13.6% of which were identified as having adverse cellular effects. This assay provides a relatively cheap and rapid approach for identifying compounds with adverse cellular effects during screening assays, potentially reducing compound rejection due to toxicity in subsequent in vitro and in vivo assays.
Insights
This study presents a new phenotypic assay for early identification of cytotoxic drug compounds during primary screening. This method improves drug discovery efficiency by detecting adverse cellular effects sooner.
Area of Science:
- Drug Discovery and Development
- Toxicology
- High-Content Imaging
Background:
- Toxicity is a primary reason for drug candidate failure.
- Current toxicological testing can be inefficient.
- Early identification of cytotoxic compounds is crucial for improving drug development success rates.
Purpose of the Study:
- To develop a novel phenotypic assay for identifying compounds with adverse cellular effects.
- To integrate cytotoxicity counter-screening into primary compound screening processes.
- To enhance the efficiency of drug discovery by early toxicity detection.
Main Methods:
- Utilized U2OS cells and the PerkinElmer Operetta high-content imaging system.
- Developed image analysis algorithms in Columbus software to detect changes in nuclear morphology, cell shape, and proliferation.
- Validated algorithms using DAPI, TOTO-3, and phosphohistone H3 staining on cells treated with known cytotoxic agents (doxorubicin, taxol, nocodazole).
Main Results:
- Successfully identified compounds with adverse cellular effects using the developed phenotypic assay.
- Screened a library of over 300 natural product-like molecules.
- Identified 13.6% of the screened compounds as having adverse cellular effects.
Conclusions:
- The novel phenotypic assay offers a cost-effective and rapid method for identifying cytotoxic compounds.
- Early detection of adverse cellular effects can significantly reduce compound rejection in later-stage in vitro and in vivo assays.
- This approach has the potential to streamline the drug discovery and development pipeline.
More Related Videos
12:22A Strategy to Identify Compounds that Affect Cell Growth and Survival in Cultured Mammalian Cells at Low-to-Moderate Throughput
Published on: September 22, 2019
08:43A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules
Published on: March 10, 2017