Performance of monocyte activation test supplemented with human serum compared to fetal bovine serum

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Marijke W. A. Molenaar-de Backer
Eelo Gitz
Miranda Dieker
Paulien Doodeman
Anja ten Brinke


The monocyte activation test (MAT) is used to detect pyrogens in pharmaceutical products and serves as replacement of the rabbit pyrogen test. The peripheral blood mononuclear cell-based MAT assay requires the addition of serum to the medium and is either performed with fetal bovine serum (FBS) or human serum (HS). Since the capacity to detect non-endotoxin pyrogens (NEPs) in a sensitive manner is an important strength of MAT compared to bacterial endotoxin test, the performance of the MAT using FBS and HS was investigated using endotoxin and several NEPs. The MAT was more sensitive for endotoxin when FBS was used, however for most NEPs the MAT was more sensitive when performed in HS. Furthermore, heat-inactivation of FBS affected the performance of the MAT for endotoxin to some extent and not for the NEPs. Interestingly, heat-inactivation of HS led to an almost complete loss of reactivity towards endotoxin, it reduced the response towards HKSA and peptidoglycan, while having minor or no effects on the responses towards R848, flagellin and Pam3CSK4. Therefore, to guarantee optimal performance of MAT heat-inactivated serum should be avoided. Moreover, product testing of a human blood-derived product in MAT using HS was beneficial since endotoxin spike recoveries were improved. This product is therefore currently batch released with the HS-based MAT assay. Overall, The HS-based MAT appears to be the first choice to replace the rabbit pyrogen test while in some cases the FBS-based MAT may be favored.

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How to Cite
Molenaar-de Backer, M. W. A., Gitz, E., Dieker, M., Doodeman, P. and ten Brinke, A. (2020) “Performance of monocyte activation test supplemented with human serum compared to fetal bovine serum”, ALTEX - Alternatives to animal experimentation. doi: 10.14573/altex.2008261.