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Related Experiment Videos

A sensitive new bioassay for tumor necrosis factor

T A Shahan1, P D Siegel, W G Sorenson

  • 1Division of Respiratory Disease Studies, National Institute for Occupational Safety and Health, Morgantown, WV 26505.

Journal of Immunological Methods
|October 14, 1994
PubMed
Summary

This study presents a highly sensitive tumor necrosis factor (TNF) bioassay using a novel cell line and fluorescence detection. The improved assay offers greater sensitivity for detecting TNF in biological fluids, aiding disease research.

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Area of Science:

  • Biotechnology
  • Immunology
  • Cell Biology

Background:

  • Tumor necrosis factor (TNF) is a key cytokine in inflammatory processes.
  • Accurate measurement of TNF in biological fluids is crucial for understanding various diseases.
  • Existing TNF bioassays have limitations in sensitivity and detection methods.

Purpose of the Study:

  • To develop an improved and highly sensitive bioassay for tumor necrosis factor (TNF).
  • To utilize a newly isolated subclone of NCTC-clone 929 cells for enhanced TNF detection.
  • To implement a novel fluorescence indicator system for improved target cell viability assessment.

Main Methods:

  • Cultivation of a hypersensitive NCTC-clone 929 subclone for TNF bioassay.
  • Employment of a fluorescence viability assay for precise detection of target cells.

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  • Optimization of assay sensitivity by evaluating different serum sources and concentrations.

    • Main Results:

    • The developed TNF bioassay achieved a limit of detection of 68 +/- 2.5 fg/ml.
    • The hypersensitive cell line demonstrated approximately 3x and 10x increased sensitivity compared to parental clones and previous methods, respectively.
    • Assay sensitivity was optimal at >= 15% serum concentration, with no significant difference between bovine fetal and horse serum sources.

    Conclusions:

    • The novel fluorescence-based TNF bioassay offers superior sensitivity for cytokine detection.
    • This improved assay provides a valuable tool for research into inflammatory diseases and related conditions.
    • Optimal cell culture conditions, including serum concentration, are critical for maintaining assay sensitivity over time.