The Monocyte Activation Test (MAT) is an in vitro assay for pyrogen testing in the biopharmaceutical or medical device industry. The MAT test principle is based on the detection of inflammatory cytokines released by human monocytes in response to exogenous endotoxins (LPS) and non-endotoxin pyrogens (NEP).
Since its first adoption into the European Pharmacopeia in 2010 (Ph. Eur. chapter 2.6.30) the MAT assay is recognized as a compendial method for pyrogen testing in a number of world regions including Europe. Here, the MAT is recommended as a replacement test for the Rabbit Pyrogen Test (RPT) (1). The MAT is further compliant to the global framework of the 3Rs principle (replacement, refinement, reduction of experimental animal tests).(2)
The PyroCell™ MAT Kit contains a human peripheral blood mononuclear cell preparation pooled from 4 healthy donors (pMAT cells), and has been specifically qualified for endotoxins and pyrogen testing with the MAT. Thereby, donor selection, cell preparation and quality testing strictly follow the requirement of the European Pharmacopeia. The optimized MAT Cell Culture Medium Supplement component of the PyroCell™ MAT Kit has been formulated with special attention to a low endotoxin content in order to ensure error-free cell stimulation conditions. The PyroCell™ MAT Kit is optimized for use with the PeliKine Compact™ human IL-6 ELISA Kit and Tool Set. All components together represent the PyroCell™ MAT Assay System, IL-6 supporting the complete testing of 3x 3 product samples according to the compendial requirements of the European Pharmacopeia. The historical LOD of the PyroCell™ MAT assay system, IL-6 is ? 0.02 EU/ml.
The MAT assay is performed in two steps: during an overnight cell culture step, a monocytic cell preparation, e.g. peripheral blood mononuclear cells (PBMC) is stimulated for release of inflammatory cytokines with a product preparation that may contain a pyrogenic contaminant. On the next day, the cell culture supernatant is analyzed for the presence of released cytokines using an ELISA assay. The color reaction is measured in an absorbance reader, e.g. the ELx808 Reader, at a wavelength of 450 nm and a reference wavelength of 540-590 nm. The pyrogenic content of the sample is then analyzed against the reference standard endotoxin (RSE) and is expressed in Endotoxin Equivalent Units (EEU/mL).
The following reagents, accessories and instuments are used to support the PyroCell™ MAT Assay System:
Step 1: Monocyte Activation
Product Catalog #
Iscove's Modified Dulbecco's Medium (IMDM) with L-Glutamine and HEPES 12-722F
USP Reference Standard Endotoxin E700
LAL Reagent Water W50-100
Pyrogen-free Dilution Tubes N207
Reagent Reservoirs 00190035
LAL Reagent Grade™ Multi-well Plates 25-340
Endotoxin-free accessories, i.e. sample container,
pipette tips, serological pipettes See Catalog
Step 2: IL-6 ELISA Assay
Product Catalog #
PeliKine Compact™ human IL-6 ELISA Assay Kit (M1916) 00253325
PeliKine Compact™ human IL-1b ELISA Assay Kit (M1934) 00253324
ELx808 Reader (with 450 and 540-590 nm filter) 25-315S
Endotoxin-free accessories, i.e. sample container,
pipette tips, serological pipettes See Catalog
(1) European Pharmacopeia
(2) Global framework 3R
Endotoxin and Pyrogen Testing Products
The next chapter in endotoxin testing... Let's write it together.
