International Validation of Pyrogen Tests Based on Cyropreserved Human Primary Blood Cells

Pyrogens as fever-inducing agents can be a major health hazard in parenterally applied drugs. For the control of these contaminants, pyrogen testing for batch release is required by pharmacopoeias. This has been done either by the in vivo rabbit pyrogen test (since Î942) or the limulus amoebocyte lysate test (LAL), since 1976. New approaches include cell-based assays employing in vitro culture of human immune cells which respond e.g. by cytokine production (IL-lβ; IL-6) upon contact with pyrogens. Six variants of these assays have been validated in a collaborative international study. The recent successful development of cryopreservation methods promises to make standardized immunoreactive primary human blood ceils available for widespread use. Furthermore, the pretesting of donors for infectious agents such as HIV or hepatitis has made it possible to develop a safe and standardised reagent for pyrogen testing. Using a total of 13 drugs, we have validated the pyrogcn test based on fresh and cryopreserved human whole blood in four laboratories. The test reached >90% sensitivity and specificity. In contrast to the LAL, tlie test was capable of detecting non-endotoxin pyrogens derived from Gram-positive bacteria or fungi.

SCHINDLER Stefanie;  SPREITZER Ingo;  LOESCHNER Bettina;  HOFFMANN Sebastian;  HENNES Kilian;  HALDER Maria;  BRUEGGER Peter;  FREY Esther;  HARTUNG Thomas;  MONTAG LESSING Thomas; 

2007-02-19

ELSEVIER SCIENCE BV

JRC36329

https://publications.jrc.ec.europa.eu/repository/handle/JRC36329,   

10.1016/j.jim.2006.07.023,