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Related Experiment Videos

A flow injection renewable surface technique for cell-based drug discovery functional assays.

P S Hodder1, J Ruzicka

  • 1Department of Chemistry, University of Washington, Seattle 98195-1700, USA. phodder@u.washington.edu

Analytical Chemistry
|March 27, 1999
PubMed
Summary

A new flow injection-renewable surface (FI-RS) method automates cell-based pharmacology assays. This technique uses microcarrier beads for reproducible measurements and improved biological response in drug discovery.

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Area of Science:

  • Pharmacology
  • Cell Biology
  • Analytical Chemistry

Background:

  • Traditional cell-based assays face challenges with diminishing biological response over repeated use.
  • Automated systems are needed for high-throughput drug screening and pharmacological analysis.

Purpose of the Study:

  • To introduce and validate a novel flow injection-renewable surface (FI-RS) technique for automated cell-based pharmacology assays.
  • To demonstrate the FI-RS technique's capability in measuring agonist-receptor interactions and quantifying pharmacologic parameters.

Main Methods:

  • Cells (Chinese hamster ovary) expressing rat muscarinic M1 receptors were attached to microcarrier beads.
  • A flow injection apparatus delivered varying concentrations of muscarinic agonists (carbachol, acetylcholine, pilocarpine).

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  • Intracellular calcium elevation was measured using fura-2 and fluorescence microscopy photometry.

    • Main Results:

    • The FI-RS technique provided reproducible control over agonist concentration gradients delivered to cells.
    • Pharmacologic parameters were quantified with results comparable to existing literature values.
    • The method successfully eliminated the diminishing biological response issue seen in traditional assays.

    Conclusions:

    • The FI-RS technique offers a robust and reproducible platform for automated cell-based pharmacology assays.
    • This method enhances assay reliability by utilizing a disposable and renewable surface (microcarrier beads).
    • The FI-RS assay shows potential as an analytical tool for discriminating agonists based on kinetic properties.