28th May 2020 Content supplied by: Merck Millipore
How Well is the Cell Line-Based MAT Suited for Pyrogen Detection?
We have demonstrated that the Mono-Mac-6 cell line used in our PyroMAT® system is well-suited for the detection of a wide range of ligands targeting all Toll-like receptors of monocytic cells. This compelling evaluation of the monocytic cells’ suitability delivers further evidence to support the use of the monocyte activation test (MAT) as a replacement for the rabbit pyrogen test, and to provide confidence when releasing parenteral drugs.
Study: Detection of various non-endotoxin pyrogens (NEPs) or TLR ligands with the PyroMAT® system.
For this study, 10 samples were used, containing Pam3CSK4, HKSA, PGN, FSL-1, Poly-IC, flagellin, imiquimod, CL075, ODN2006 or MDP. The appropriate concentration range of each ligand was determined by dose-screening using one batch of PyroMAT® cells, followed by verification of the reaction using three further batches.
Determination of limit of detection (LOD) for the different NEPs
The different samples were used to determine the LOD of the MAT for each ligand. For this, at least four dilutions of every NEP were run individually in the assay. The pyrogenicity of each concentration, measured as endotoxin equivalent units (EEU) per millilitre, is shown below in “Graphs”.
All the NEPs tested were detected, with all values over the cut-off of the respective assay. The contaminant samples with MDP and ODN2006 could be detected but not quantified, as the calculated values were below the validated limit of quantification, 0.05 EU/mL.
Date Published: 28th May 2020
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