The meningococcal C conjugate vaccine (MenCC) is an interesting model with which to testthe efficacy of the Monocyte Activation Test (MAT) as an alternative method of pyrogen testing in thequality control of vaccines. The MenCC that has been produced by Bio-Manguinhos in Brazil is in the finaldevelopment stage, and, as recommended in the guidelines for MenCC production, its pyrogen contentmust be determined by using the Limulus Amoebocyte Lysate (LAL) assay and the Rabbit Pyrogen Test (RPT).This represents an ideal opportunity to compare LAL and RPT data with data obtained by using a MATsystem with cryopreserved whole blood and IL-6/IL-1as marker readouts. In order to assess thecompatibility of the MAT with MenCC, endotoxin and non-endotoxin pyrogen content was quantified byusing MenCC samples spiked with lipopolysaccharide (LPS), lipoteichoic acid or zymosan standards. Thepresence of the aluminium-based adjuvant interfered with the MAT, increasing the readout of IL-1in LPS-spiked MenCC batches. This infringed the product-specific validation criteria of the test, and led to IL-6being chosen as the more suitable marker readout. No pyrogenic contaminants were identified in theMenCC batches tested, demonstrating consistency among the different systems (MAT, RPT and the LALassay). In conclusion, the introduction of the MAT during MenCC development could contribute to theelimination of animal tests post-licensing, ensuring human protection based on an effective non-animalbased method of quality control.