Cellular Bioluminescence Based Assay: A High Throughput Method for Combinatorial Drug Screening
Encyclopedia of Experiments
Cancer Research
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Encyclopedia of Experiments Cancer Research
Cellular Bioluminescence Based Assay: A High Throughput Method for Combinatorial Drug Screening

Cellular Bioluminescence Based Assay: A High Throughput Method for Combinatorial Drug Screening

Transcript

Cellular bioluminescence is a highly sensitive and rapid approach for large-scale screening of anti-cancer drugs.

This technique uses tumor cells expressing luciferase - an enzyme that catalyzes a light-producing chemical reaction, causing the host cells to glow in the dark. The intensity of the emitted luminescence is directly proportional to the live cell density.

To begin, seed luciferase-expressing cells in an optically clear culture plate coated with an extracellular matrix or ECM and incubate.

During incubation, the ECM promotes the attachment of cells to the culture well.

Next, take concentrated stocks of the drug combination and add different volumes of culture media to prepare serial dilution of the drug mixture.

Now, remove the culture medium from the adhered cells. Add different dilutions of the drug combination and incubate.

During incubation, the drug combination acts synergistically on cellular targets.

Now, remove the drug-containing medium and add fresh medium containing luciferin.

Luciferin diffuses across the membrane and enters the cells. The luciferases expressed by the cells use cellular ATP to oxidize luciferin and emit light signals.

Quantify the cellular bioluminescence under an imaging system.

A decrease in light emission with increased drug concentration indicates reduced cell viability. This correlates to the drug's antiproliferative effect on the target cells.

Key Terms and Concepts

  • High-throughput screening - A method for large-scale testing of substances, often in drug discovery.
  • Antiproliferative drugs - Medications aimed at inhibiting cell growth or division.
  • Bioluminescence test - A technique to detect and quantify specific substances based on their ability to emit light.
  • Luciferin-Luciferase - A system in which the luciferase enzyme catalyzes the oxidation of luciferin, producing light.
  • Drug combination screening - The process of testing various drug mixtures to establish their combined effect on an organism.

Scientific Background

  • Introduce bioluminescence - A natural phenomenon where light is produced and emitted by a living organism (e.g., luciferase).
  • Key Concepts - The luciferase enzyme oxidizes luciferin to produce light, and this can be used in drug testing (e.g., bioluminescence assay).
  • Underlying Mechanisms - Light emission decreases with an increase in drug concentration, indicating the antiproliferative effect of the drug on cells (e.g., high-throughput screening).
  • Connect to Experiment - This bioluminescence model is employed in cellular testing to screen potential antiproliferative drugs.

Questions This Video Helps You Answer

  • What is 'bioluminescence' and how is it used in 'high-throughput screening' applications?
  • How does luciferase react with luciferin in 'bioluminescence assays'?
  • What changes in bioluminescence indicate in 'drug combination screening'?

Applications and Relevance

  • Practical Applications - High-throughput screening is crucial in drug discovery and pharmacological research (e.g., antiproliferative drugs).
  • Industry Impact - Drug development, medical diagnostics, and biotechnology sectors are greatly impacted by these techniques (e.g., bioluminescence assays).
  • Societal Importance - These assays contribute towards finding cures for serious health conditions, improving public health (e.g., bioluminescence tests).
  • Link to Scientific Advancements - Advancements in bioluminescence techniques have accelerated the drug discovery process.

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