The method transfer and validation of freeze-dried rabies vaccine pyrogen detection in HL-60 monocyte activation assay
- Author:
Chen CHEN
1
;
Yuan ZHANG
1
;
Hua GAO
1
;
Yan-Lin WU
1
;
Can WANG
2
Author Information
- Collective Name:National Institutes for Food and Drug Control
- Publication Type:Journal Article
- Keywords: freeze-dried rabies vaccine for human use; human promyelocyte leukemia HL-60 cells; inter- leukin 6( IL-6); method transfer; method validation; mon¬ocyte activation experiment; pyrogen detection
- From: Chinese Pharmacological Bulletin 2022;38(10):1586-1590
- CountryChina
- Language:Chinese
- Abstract: Aim To study the application of HL-60 monocyte activation test in the pyrogen detection of freeze-dried rabies vaccine for human use.Methods The established HL-60 mononuclear cell activation test (MAT) was transferred between laboratories and the method was verified; referring to the interference test in the photometric method of the bacterial endotoxin test method of the Chinese Pharmacopoeia, HL-60- IL- 6 MAT was used to detect the recovery and pyrogen content of 13 batches of freeze-dried rabies vaccine for human use.Results The linearity of the amount of 1L-6 secreted by HL-60 cells, which stimulated by dif¬ferent concentrations of endotoxin standards was above 0.95; the calculated minimum detection limit was not more than 0.125 EU • mL"1 ; the recovery experiment with a solution containing 0.5 and 1.0 EU • mL"1 of endotoxin was performed to cheek the accuracy of the method.HL-60-IL-6 was used to detect 13 hatches of Freeze-dried rahies vaccine for human use, and the re¬covery of endotoxin was between 50 % to 200%.It was consistent that HL-60-IL-6 with pyrogens and en¬dotoxin test for 4 batches of freeze-dried rabies vaccine for human use which pyrogens and endotoxin test failed and the 3 batches of water for injection.Conclusion The HL-60 MAT using IL-6 as a detection indicator is suitable for the detection of pyrogenic substances in freeze-dried rabies vaccine for human use.
