More Flexible MAT Assay Design by Taking Advantage of Reporter Assay-Based MAT: Poster presented at PDA Microbiology Conference 2025.

Abstract

We have developed a next-generation MAT kit based on a reporter assay. In this presentation, we report on the potential for a more flexible MAT test design using our new reagent.In conventional MAT, inflammatory cytokines are typically detected using ELISA as an indicator. In contrast, the LumiMAT pyrogen detection kit detects the transcriptional activity of NF-κB, a transcription factor that induces the expression of inflammatory cytokines, using a luciferase reporter assay. Compared to ELISA-based MAT, the advantages of the reporter assay-based MAT include a significant reduction in cell incubation time due to increased sensitivity (from 24 hours to 3 hours) and the removal of labor-intensive steps (several hours for ELISA to a few minutes for the addition of luminescent substrates).For practical application in pyrogen testing, we conducted preliminary tests in accordance with EP 2.6.30. guidelines on several medicinal products subject to Rabbit Pyrogen Test and confirmed that the results met the requirements specified in the pharmacopoeia. Additionally, our method allows for reduced coefficient of variation, enabling the reduction in the number of replicates (n=4 to n=3) and increasing throughput (from a 96-well plate to a 384-well plate). These features demonstrate the potential for designing more cost-effective test protocols.