Development and validation of a novel monocyte activation test with secreted luciferase-expressing cells

Clicks: 1
ID: 315912
2026
Free PDF
Article Quality & Performance Metrics
Overall Quality
Not rated
Combines reader engagement with the AI quality analysis. This article has not been analysed, so there is no overall score — reader engagement is measured and shown alongside.
AI Quality Assessment
Not analyzed
Readership in this journal

Ranked #24 of 36 articles by views in The Journal of Biochemistry

Most read Least read

Bar heights use a square-root scale.

Mint this article as an NFT
Not yet minted

Create a permanent, verifiable on-chain record of this article on the Scimatic Network. The NFT is held in your Journament account, and you can withdraw it to your own wallet at any time.

5 SUSD one-off · no wallet required
Sign in to mint
Abstract
Pyrogen testing is essential for ensuring pharmaceutical product safety; however, current methods have limitations. The Bacterial Endotoxins Test cannot detect non-endotoxin pyrogens (NEPs), whereas the Rabbit Pyrogen Test was phased out for animal welfare reasons. Monocyte activation tests (MAT), using human whole blood samples or peripheral blood mononuclear cells, are effective but suffer from donor variability, biosafety concerns, and instability. We aimed to develop a sensitive, rapid, and simple alternative pyrogen test by devising a luciferase-based MAT using the Mono-Mac-6 (MM6) cell line. MM6 cells showed higher cytokine responses to endotoxin than THP-1 cells and exhibited comparable or greater sensitivity than human peripheral blood. Introducing an NF-κB-driven NanoLuc® reporter enabled endotoxin detection within 3 h; however, rapid degradation of intracellular luciferase was noted. To overcome this degradation issue, we generated MM6 cells expressing secreted NanoLuc (secNluc), allowing for stable detection between 3 and 12 h after stimulation. The MM6/NFκB-secNluc cells helped detect various pyrogens, including Pam3CSK4, peptidoglycan, poly(I:C), lipopolysaccharide, flagellin, macrophage-activating lipopeptide-2, and resiquimod. Adding recombinant human lipopolysaccharide-binding protein enabled establishing a serum-free MAT system, eliminating serum-derived endotoxin contamination. This reporter-based MAT provides a robust platform for rapid endotoxin and NEP detection, offering an improved alternative to conventional MATs.
Reference Key
openalex_W7163509081 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Teruaki Oku, Yusuke Ando, Tomohisa Nanao, Yutaka Kikuchi, Yukiko Hara‐Kudo
Journal The Journal of Biochemistry
Year 2026
DOI
10.1093/jb/mvag038
URL
https://doi.org/10.1093/jb/mvag038
Keywords Keywords not found

Citations

No citations found. To add a citation, contact the admin at info@scimatic.org

Comments

No comments yet. Be the first to comment on this article.