Development and validation of a novel monocyte activation test with secreted luciferase-expressing cells - PubMed
3 hours ago
- #luciferase reporter
- #pyrogen testing
- #monocyte activation test
- A novel monocyte activation test (MAT) using luciferase-expressing cells was developed to address limitations in current pyrogen tests.
- The test uses the MM6 cell line, which showed higher cytokine responses to endotoxin compared to THP-1 cells and comparable sensitivity to human peripheral blood.
- An NF-κB-driven secreted NanoLuc reporter allowed for stable detection of pyrogens within 3 to 12 hours, overcoming rapid degradation issues with intracellular luciferase.
- The system detected various pyrogens, including endotoxins and non-endotoxin pyrogens like Pam3CSK4 and peptidoglycan.
- Adding recombinant human lipopolysaccharide-binding protein enabled a serum-free MAT system, eliminating serum-derived endotoxin contamination.
- This reporter-based MAT offers a robust, rapid, and simple alternative to conventional methods, reducing donor variability and biosafety concerns.