The Monocyte Activation Test (MAT) is an in vitro assay for pyrogen testing in pharmaceutical and medical device industries. The principle of the Monocyte Activation Test is based on the detection of inflammatory cytokines released by human monocytes in response to exogenous bacterial endotoxin (LPS) and non-endotoxin pyrogens (NEP). Since its first adoption by the European Pharmacopeia in 2010 (Ph. Eur. chapter 2.6.30) the Monocyte Activation Test is recognized as a compendial method for pyrogen testing in several world regions including Europe. Here, the Monocyte Activation Test is recommended as a replacement test for the Rabbit Pyrogen Test (RPT) (1). The Monocyte Activation Test is compliant to the EU Directive 2010/63/EU on the protection of experimental animals and the global framework of the 3Rs principle (replacement, refinement, reduction of experimental animal tests). (2) The PyroCell® Monocyte Activation Test – Human Serum Kit (PyroCell® MAT HS Kit) contains a peripheral blood mononuclear cell preparation pooled from 4 human blood donations (pMAT cells), and has been specifically qualified for endotoxin and pyrogen testing with the Monocyte Activation Test. Thereby, donor selection, cell preparation and quality testing strictly follow the requirement of the European Pharmacopeia. The optimized Human Serum Supplement component of the PyroCell® Monocyte Activation Test – Human Serum Kit has been formulated with special attention to a low endotoxin content in order to ensure error-free cell stimulation conditions. The PyroCell® Monocyte Activation Test – Human Serum Kit is optimized for use with the PeliKine compact human IL-6 ELISA Kit. All components together represent the PyroCell® MAT HS System supporting the complete testing of 3x 2 product samples according to the requirements of the European Pharmacopeia. The historical LOD of the The PyroCell® Monocyte Activation Test – Human Serum system is ≤ 0.08 EU/mL. The PyroCell® Monocyte Activation Test is performed in two steps: during an overnight cell culture step, the peripheral blood mononuclear cells (PBMC) preparation is stimulated for release of inflammatory cytokines with a product sample to be tested for the presence of pyrogens. On the next day, the cell culture supernatant is analyzed for released IL-6 cytokines using an human IL-6 ELISA assay. The color reaction is measured in an absorbance reader at a wavelength of 450 nm and a reference wavelength of 540-590 nm. The pyrogen 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 instruments support the use of the PyroCell® MAT HS Kit : 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 190035 LAL Reagent GradeTM 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 00253325 ELx808 Reader (with 450 and 540-590 nm filter) 25-315S (1) European Pharmacopeia (2) Global framework 3R
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