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Related Experiment Videos

High throughput screening technologies for direct cyclic AMP measurement.

Daniela Gabriel1, Magali Vernier, Martin J Pfeifer

  • 1Novartis Pharma AG, Lead Discovery Center, Basel, Switzerland. daniela.gabriel@pharma.novartis.com

Assay and Drug Development Technologies
|April 20, 2004
PubMed
Summary
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This study compared five cAMP detection technologies for high-throughput screening (HTS). AlphaScreen and HTRF demonstrated superior sensitivity and HTS compatibility, making them ideal for G(s)-coupled antagonist screening.

Area of Science:

  • Biochemistry
  • Assay Development
  • Pharmacology

Background:

  • Cyclic adenosine monophosphate (cAMP) is a crucial second messenger involved in various cellular signaling pathways.
  • Accurate and efficient detection of cAMP is essential for drug discovery and understanding cellular functions.
  • High-throughput screening (HTS) requires sensitive, robust, and feasible assay technologies.

Purpose of the Study:

  • To compare the performance of five different cAMP detection technologies for HTS.
  • To evaluate sensitivity, robustness, and feasibility of each method.
  • To identify the most suitable technologies for specific screening applications, such as G(s)-coupled antagonist screens.

Main Methods:

  • Nonhomogeneous DELFIA (Dissociation Enhanced Lanthanide Fluoroimmunoassay)

Related Experiment Videos

  • Homogeneous time-resolved fluorescence (HTRF)
  • Homogeneous luminescent singlet oxygen channeling (AlphaScreen)
  • Fluorescence Polarization (FP)
  • High-affinity enzyme complementation

    • Main Results:

    • DELFIA exhibited the highest sensitivity among the tested methods.
    • AlphaScreen and HTRF offered a balance of high sensitivity, broad dynamic range, and simplified workflows with minimal reagent addition.
    • For G(s)-coupled antagonist screening, HTRF and AlphaScreen were identified as the most sensitive and HTS-compatible techniques.

    Conclusions:

    • AlphaScreen and HTRF are highly suitable for HTS of cAMP modulation, particularly for antagonist screening.
    • The choice of cAMP detection technology should consider the specific requirements of the screening assay, including sensitivity and compatibility with HTS platforms.
    • Further optimization of these homogeneous assays can enhance their utility in drug discovery.