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

• DOI: 10.1038/s41392-024-01744-0
• Publication Date: 2024-02-19T01:01:55Z
• 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
• Keywords: Zymosan

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