破伤风类毒素和白喉类毒素原液的质量分析

Quality analysis of tetanus toxoid and diphtheria toxoid bulks

目的 分析破伤风类毒素（tetanus toxoid,TT）和白喉类毒素（diphtheria toxoid,DT）原液的细菌内毒素含量、单体与多聚体之比以及游离氨基数.方法 取TT和DT原液各10批,用凝胶法、高效液相色谱法及三硝基苯磺酸法分别检测原液的细菌内毒素、单体和多聚体以及游离氨基.结果 各10批TT、DT原液的细菌内毒素含量均值分别为1.14和0.35 EU/Lf.TT原液的单体平均含量为72.1％,是多聚体的3倍;DT原液的单体平均含量为88.9％,是多聚体的8倍.脱毒后,每个TT蛋白分子含游离氨基数均值为43.5,比破伤风毒素减少约60％;每个DT蛋白分子含游离氨基数均值为9.9,比白喉毒素减少约75％.10批TT、DT的单体与多聚体含量之比以及游离氨基数的批间一致性较好（变异系数：1.59％～16.26％）.结论 细菌内毒素含量、单体与多聚体之比以及游离氨基数检测指标可用于对TT、DT生产过程的控制以及批间一致性的监控.

Objective To analyze endotoxin contents,monomer/polymer ratio,and number of free amino groups in tetanus toxoid （TT） and diphtheria toxoid （DT） bulks.Methods Ten batches each of TT and DT bulks were taken.Gel-clot test,high performance liquid chromatography,and trinitro benzene sulfonic acid method were used to detect bacterial endotoxin,monomer/polymer,and free amino groups,respectively.Results The average endotoxin contents of TT and DT bulks were 1.14 and 0.35 EU/Lf,respectively.TT monomer content averaged 72.1%,3 times of polymer content.DT monomer content averaged 88.9%,8 times of polymer content.Average number of free amino groups per protein molecule was 43.5 in TT and 9.9 in DT.Compared with their respective toxins,free amino group number after detoxification decreased by approximately 60% in TT and 75% in DT.In both TT and DT,monomer/polymer ratio and free amino group contents showed good batch consistency （coefficient of variation：1.59%-16.26%）.Conclusion Bacterial endotoxin content,monomer / polymer ratio,and free amino group number can be used as detection indicators in controlling TT and DT production process and monitoring the batch consistency of TT and DT.