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Cytokine release assay


The aim of this study is to measure the cytokine release (selected cytokines or cytokine panel) as part of the investigation of immunological response of an organism to a test item.

The basis of the quantification of the cytokine production is ELISA (Please find a general description of method establishment and validation of the method at ELISA development and validation section). For quantification of multiple cytokine panel (especially in case of small volume of biological matrix), Luminex measurement can be applied using commercially available custom cytokine panel assays. These measurements can be part of a toxicity study or clinical trial.


Usually two species: one rodent and one non-rodent.


Establishment/transfer the method, selection of suitable reagents, setting up the validation targets and acceptance criteria.

Performance of validation testing the following parameters:

Quantification range (LLOQ, ULOQ)

  • Accuracy at different level
  • Precision at different level
  • Selectivity and matrix effect
  • Sensitivity
  • Spiking recovery
  • Dilution linearity
  • Stability of target molecules in the matrix


Concentration of cytokine (or cytokine panel) in the biological matrix.

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