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A novel alternative for pyrogen detection based on a transgenic cell line

Authors :
Qing He
Chuan-Fei Yu
Gang Wu
Kai-Qin Wang
Yong-Bo Ni
Xiao Guo
Zhi-Hao Fu
Lan Wang
De-Jiang Tan
Hua Gao
Can Wang
Gang Chen
Xu-Hong Chen
Bo Chen
Jun-Zhi Wang
Source :
Signal Transduction and Targeted Therapy, Vol 9, Iss 1, Pp 1-9 (2024)
Publication Year :
2024
Publisher :
Nature Publishing Group, 2024.

Abstract

Abstract Pyrogen, often as a contaminant, is a key indicator affecting the safety of almost all parenteral drugs (including biologicals, chemicals, traditional Chinese medicines and medical devices). It has become a goal to completely replace the in vivo rabbit pyrogen test by using the in vitro pyrogen test based on the promoted ‘reduction, replacement and refinement’ principle, which has been highly considered by regulatory agencies from different countries. We used NF-κB, a central signalling molecule mediating inflammatory responses, as a pyrogenic marker and the monocyte line THP-1 transfected with a luciferase reporter gene regulated by NF-κB as an in vitro model to detect pyrogens by measuring the intensity of a fluorescence signal. Here, we show that this test can quantitatively and sensitively detect endotoxin (lipopolysaccharide from different strains) and nonendotoxin (lipoteichoic acid, zymosan, peptidoglycan, lectin and glucan), has good stability in terms of NF-κB activity and cell phenotypes at 39 cell passages and can be applied to detect pyrogens in biologicals (group A & C meningococcal polysaccharide vaccine; basiliximab; rabies vaccine (Vero cells) for human use, freeze-dried; Japanese encephalitis vaccine (Vero cells), inactivated; insulin aspart injection; human albumin; recombinant human erythropoietin injection (CHO Cell)). The within-laboratory reproducibility of the test in three independent laboratories was 85%, 80% and 80% and the interlaboratory reproducibility among laboratories was 83.3%, 95.6% and 86.7%. The sensitivity (true positive rate) and specificity (true negative rate) of the test were 89.9% and 90.9%, respectively. In summary, the test provides a novel alternative for pyrogen detection.

Details

Language :
English
ISSN :
20593635
Volume :
9
Issue :
1
Database :
Directory of Open Access Journals
Journal :
Signal Transduction and Targeted Therapy
Publication Type :
Academic Journal
Accession number :
edsdoj.3d4236daa46a42f883ceb75d9255815f
Document Type :
article
Full Text :
https://doi.org/10.1038/s41392-024-01744-0