Application of Monocyte Activation Test in the Pyrogen Detection of Vaccine / 中国药师

ABSTRACT

Objective:To study the application of HL-60 monocyte activation test in the pyrogen detection of vaccines. Methods:HL-60 cells were incubated with the standard pyrogen(endotoxin,zymosan and lipoteichoic acid)solution and the sample solution at different concentrations,and then IL-6 level was detected using ELISA kit. According to the concentration of standard pyrogen solution and the level of IL-6,the standard curve and linear relationship were obtained. Based on the IL-6 value from the sample solution,the pyrogen content of the sample was calculated. The standard pyrogen solution at middle concentration of the standard curve was used as the interference sample,and the interference assay was performed referring to the bacterial endotoxin test in Chinese pharmacopoeia. The pyrogen recovery rate and the content were detected using HL-60 / IL-6 assay in seven kinds of vaccines such as influenza vaccine(Split Virion),inactivated;hepatitis A(live)vaccine,freeze-dried;rabies vaccine(Vero Cell)for human use,freeze-dried;lyophilized human measles and rubella combined vaccine,live;measles and mumps combined vaccine, live; haemorrhagic fever with renal syndrome bivalent vaccine, inactived; group A and C meningococcal polysaccharide vaccine. Results:① The linear relationship between the pyrogen concentration and the IL-6 level was good,and R2 value was 0. 996,0. 993 and 0. 997,respectively. ② The recovery rate of pyrogen in the above vaccines ranged from 50% to 200% . ③ The endotoxin concentration of 10 batches of group A and C meningococcal polysaccharide vaccine was detected using HL-60 / IL-6 assay,and the average value of 6 batches from factory A was 134. 73 EU·ml-1 ,and that of 4 batches from factory B was 40. 75 EU·ml -1 . The pyrogen in the other vaccines was all negative. Conclusion:The HL-60 / IL-6 assay is suitable for the pyrogen detection of the seven kinds of vaccines.