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EU legislation now requires replacement of in vivo pyrogen tests with validated in vitro methods. With the Monocyte Activation Test (MAT), Solvias offers the only real alternative to animal testing and a more relevant prediction of human response to all pyrogens.
Testing to detect pyrogen contamination is an essential safety measure for all parenterally administered pharmaceutical products and medical devices. Pyrogens, a chemically heterogeneous group of fever-inducing compounds, can be divided into two distinct categories: endotoxin pyrogens (LPS) and non-endotoxin pyrogens (NEP). Contamination by pyrogens can result in symptoms ranging from slight fever to shock and death.
Testing procedures for the detection of pyrogens are included in every pharmacopoeia. Until recently, the only universally accepted method with the required specificity for all pyrogens was the rabbit pyrogen test (RPT), which measures the fever response of rabbits after injection of the test preparation. Alternative pyrogen tests, such as the BET (bacterial endotoxin test) and LAL (limulus amoebocytelsylate) test, have been developed to replace the RPT but their use has been limited as they are unable to detect pyrogens from sources other than endotoxins from gram-negative bacteria. Therefore, these alternative tests cannot be used for emerging cell and gene therapy products or biopharmaceuticals produced in organisms other than E. coli. Due to the complexity of today's medicinal products, an assay able to detect all pyrogens is required.
That is why Solvias developed the monocyte activation test (MAT). The MAT is a completely human test system and the only real alternative to the rabbit pyrogen test. The assay brings human monocytes into contact with the test material and evaluates the cell response by measuring the cytokine Interleukin-6. As an in vitro method, however, it must be validated for each individual product, therefore requiring additional steps as compared to the simple test by injection into test animals.
Three different formats of the MAT are internationally validated: blood based, peripheral blood mononuclear cell (PBMC) based, and cell-line based. The MAT with a cell-line is able to detect all human pyrogens and offers all the stability advantages of a cell-line based assay. It is thus ideally suited to QC batch release testing.
The MAT should now be used instead of animal testing in accordance with the EU legislative framework as stated in the Ph. Eur. 2.6.30. According to European legislation on the protection of animals for scientific use, once a validated in vitro method exists, in vivo animal tests must be replaced (Directive 2010/63/EU). Furthermore, as release testing for the European Union must be carried out within the EU (Directive 2001/83/EC, article 51), the MAT must be integrated into testing procedures within a very short time frame.
In keeping with this legislation, the Paul-Ehrlich-Institute in Germany announced in March 2019 that the German state authorities would no longer accept the use of the RPT. Other EU member states are expected to follow, thereby eliminating pyrogen testing on animals in Europe.
Solvias brings more than ten years of experience with the assay to offer expert development of testing procedures for all medicinal products concerned by the ban. Simplified by the use of ready-to-use cells from a monocytic cell line, the MAT can be run in any laboratory, delivering robust, accurate and precise results for pyrogen detection. Solvias provides full development and testing services according to customer requirements.
The assay is also available as the PyroMAT® kit from Merck Millipore.