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Published on: March 10, 2017
Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities
James Inglese1, Douglas S Auld, Ajit Jadhav
1NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892-3370, USA. jinglese@mail.nih.gov
Summary
Quantitative high-throughput screening (qHTS) generates detailed concentration-response data for large compound libraries, improving accuracy and accelerating drug discovery by identifying reliable chemical probes and leads.
Area of Science:
- Chemical Biology
- Drug Discovery
- Pharmacology
Background:
- High-throughput screening (HTS) is crucial for identifying modulators of molecular targets in pharmaceutical development.
- Traditional single-concentration HTS methods suffer from high rates of false positives and negatives, necessitating extensive follow-up.
- Comprehensive profiling of large chemical libraries requires efficient generation of pharmacological data and reliable activity assessment.
Purpose of the Study:
- To develop and validate a quantitative high-throughput screening (qHTS) paradigm for comprehensive profiling of large chemical libraries.
- To demonstrate the feasibility of generating concentration-response curves for thousands of compounds in a single experiment.
- To enable rapid identification of active compounds and elucidation of structure-activity relationships directly from primary screening.
Main Methods:
- Developed quantitative high-throughput screening (qHTS) to generate concentration-response curves for over 60,000 compounds.
- Applied qHTS to the enzyme pyruvate kinase as a model system.
- Classified concentration-response curves to identify activators and inhibitors with varying potencies and efficacies.
Main Results:
- qHTS precisely generated concentration-response data, proving robust against sample preparation variations.
- The method identified pyruvate kinase modulators with diverse potencies and efficacies, revealing structure-activity relationships.
- Comparison with traditional HTS showed a high prevalence of false negatives in single-concentration screens.
Conclusions:
- qHTS enables accurate and efficient profiling of large chemical libraries (>10^5 compounds).
- The rich data generated by qHTS facilitates immediate mining for reliable biological activities.
- qHTS serves as a powerful platform for chemical genomics and accelerates lead identification in drug discovery.
