无细胞百日咳疫苗效力国家参考品的建立

Establishment of National Reference for Potency Assay of Acellular Pertussis Vaccine

采用罐培养百日咳CS菌株，培养物经硫酸铵两段盐析、浸提后，再经蔗糖密度梯度离心，收集以FHA．PT为主的百日咳菌保护性抗原，经福尔马林解毒，制备成原液。该原液按照中国和日本的生物制品规程检定合格后，加入保护剂，稀释至18μgPN／ml，分装冻干制备成无细胞百日咳疫苗效力参考品。以JNIH3和JNIH－6参考品为标准，按照日本规程的小鼠脑腔攻击法标定其效力单位。根据14次效力试验所得的结果和日本NIH标定的结果，该批疫苗效力国家参考品的效力单位定为15PIU／ml。

The active components of Bordetella pertussis bacteria were extracted fromthe CS strain cultured in fermenter by fractional salting out with ammonium sulfate anddensity gradient centrifugation with sucrose. After being detoxified with formalin, the extractwas used as the stock solution of acellular pertussis vaccine. The stock solution passed thedetermination according to Chinese and Japanese Requirements and was diluted to a finalconcentration of 18μg PN/ml after protective agent was added in, then lyophilized as acandidate of national reference for pertussis potency test. The potency unit of the candidate wasassigned as 15PIU/ml by murine intracranial challeng according to Japanese Requirements,using JNIH-3 and JNIH-6 references as standards.