Pyrogenicity

In vitro Pyrogen Test

Pyrogens are fever causing agents like components of microorganisms and material surfaces. In parenterally applied drugs and medical devices in contact with the human body they can be a major health hazard. Therefore regulatory requirements demand for testing their potential pyrogenicity. Conventional methods are afflicted with some disadvantages. These disadvantages can be circumvented by the in vitro pyrogen test on the basis of cytokine release of activated monocytes after contact to pyrogens. Pyrogen test: Monocyte activation test (MAT) with human blood The in vitro pyrogen test is an internationally validated method which measures the release of cytokines from human blood cells exposed to pyrogenic compounds or material surfaces in vitro (EP 6.7, 2.6.30).

Advantages of the in vitro (EP 6.7, 2.6.30) pyrogen test:
• Detection of a wide range of pyrogens (endotoxins and non-endotoxins).
• Use of human blood in vitro is most relevant to the physiological in vivo situation.
• Replacement of the in vivo rabbit pyrogen test.
• The test is internationally validated.

Method
• Step 1:
  Incubation of sample or sample extract with human blood. Monocytes exposed to Pyrogens will release cytokines e.g. Interleukin-1ß (IL-1ß).
• Step 2:
  Analysis of IL-1ß concentration in blood by ELISA technology. Determination to pyrogenicity through a threshold concentration.