The Monocyte Activation Test (MAT) is an in vitro assay for pyrogen testing in pharmaceutical and medical device industries. The Monocyte Activation Test principle is based on the detection of pro-inflammatory cytokines released by human monocytes in response to exogenous endotoxins (LPS) and non-endotoxin pyrogens (NEP). The Monocyte Activation Test has been adopted by the European Pharmacopeia in 2010 (Ph. Eur. chapter 2.6.30) Today, the Monocyte Activation Test is MAT recognized as a compendial method for pyrogen testing in several world regions including Europe. Here, the Monocyte Activation Test will be fully replacing the Rabbit Pyrogen Test (RPT) as of 2026 (1). The Monocyte Activation Test is further compliant to the EU Directive 2010/63/EU on the protection of experimental animals and the global framework of the 3Rs (replacement, refinement, reduction of experimental animal tests). (2) Each PyroCell® Monocyte Activation Test is performed in two steps: during an overnight cell culture step, the monocytic cell preparation consisting of pooled peripheral blood mononuclear cells (PBMC, pMAT) 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 IL-6 cytokines using the Pelikine Human IL-6 Rapid ELISA assay. The color reaction is measured using a multimode reader, e.g the Nebula® Absorbance Reader or the Nebula® Multimode Reader at a wavelength of 450nm and a reference wavelength of 540-590nm or alternatively, an absorbance reader, e.g. the ELx808TM Reader, at the same wavelength. 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 PyroCell® Monocyte Activation Test Rapid System (PyroCell® MAT Rapid System) is an all-inclusive kit containing all necessary reagents for the Monocyte Activation Test: the PyroCell® Monocyte Activation Test Kit (PyroCell® MAT Kit) that contains the cryopreserved, human PBMC preparation pooled from 4 healthy human blood donations (pMAT Cells) along with an optimized MAT Culture Medium Supplement, and the Pelikine Human IL-6 ELISA Rapid Kit. that contains all reagents and buffer concentrates to conduct the human IL-6 ELISA. pMAT Cells are specifically qualified for pyrogen detection with the Monocyte Activation Test, e.g. are certified for their ability to detect endotoxin and non-endotoxin pyrogens. Thereby, donor selection, cell preparation and quality testing strictly follow the compendial requirements outlined in the Ph. Eur. Chapter 2.6.30. The MAT Culture Medium supplement is formulated with special attention to low endotoxin content in order to ensure error-free cell stimulation conditions. The PyroCell® MAT Rapid Kit has been validated for use with the PeliKine Human IL-6 ELISA Rapid Kit for sensitive detection of the released pro-inflammatory cytokine interleukin 6 (IL-6). The new Pelikine Human IL-6 ELISA Rapid Kit is composed of an ELISA detection kit and buffer concentrates ensuring rapid and highly sensitive detection of the human pro-inflammatory cytokine interleukin 6 (IL-6). Thereby, the Pelikine Human IL-6 ELISA Rapid Kit is using the same reagents proven for specific human IL-6 detection for the last 3 decades with the renowned Pelikine compact human lL-6 ELISA kit. It further contains pre-coated ELISA plates along with an optimized, rapid protocol. The PyroCell® Monocyte Activation Test Rapid System provides the optimal tool for specific detection of the human IL-6 cytokine released by human monocytes during exposure to endotoxins or non-endotoxin pyrogens. All components together support the complete testing of 3x 96-well microtiter plates (288 tests) or up to 9 product samples according to the compendial requirements for the Monocyte Activation Test. The historical LOD of the PyroCell® Monocyte Activation Test Rapid System is ≤0.02 EU/ml. (1) European Pharmacopeia (2) Global framework 3R
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