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

作　　者：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 

机构地区：[1]National Institutes for Food and Drug Control,Beijing,China [2]School of Life Science and Biopharmaceutics,Shenyang Pharmaceutical University,Shenyang,China [3]Shanghai Institute for Food and Drug Control,Shanghai,China [4]KeyMed Biosciences Inc.,Chengdu,China

出　　处：《Signal Transduction and Targeted Therapy》2024年第3期1221-1229,共9页信号转导与靶向治疗（英文）

基　　金：supported by the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences(research on quality evaluation and standardisation of innovative vaccines,2021-I2M-5-005);the National Key Laboratory of Drug Regulatory Sciences Fund(research on the alternative test for pyrogen detection based on transgenic cells,2023-NIFDC-28).

摘　　要：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.

关 键 词：PYROGEN RABBIT replace 

分 类 号：R95[医药卫生—药学]