第三批吸附白喉类毒素国家标准品的制备和标定

Preparation and calibration of the third batch of National Standard for Diphtheria Toxoid

目的建立第三批吸附白喉类毒素国家标准品,用于效价测定。方法选用白喉疫苗生产菌株(PW8,CMCC38007),经复苏传代和产毒培养后,收获白喉毒素,采用先精制后脱毒的方法获得精制白喉类毒素原液,再加入氢氧化铝吸附后分装冻干,作为备选品。按《中国药典》三部(2010版)要求对原液和冻干品进行检定。以WHO国际标准品(07/216)和中国国家标准品(002)为标准,采用小鼠Vero细胞法在3个实验室进行协作标定。结果该批标准品的各项检测指标均符合国家标准品的规定;经协作标定,获得的20个效价单位标定结果合并统计后几何均值为289.3 IU/ml,95%可信区间为276.0~303.2;同时用WHO国际标准品(07/216)和中国国家标准品(002)进行标定比较,两者差异无统计学意义(P>0.05);标准品效价稳定性考核结果证明,我国白喉类毒素国家标准品在保存期内效价稳定。结论所制备的标准品各项指标均符合要求,其效价定为289.3 IU/支,可作为第三批吸附白喉类毒素国家标准品用于效价测定。

Objective To establish the third batch of National Standard for Diphtheria Toxoid for potency assay.Methods The production strains of diphtheria vaccine, PW8 and CMCC 38007, were resuscitated and cultured for producing toxin. The bulk of refined diphtheria toxoid was obtained after toxin extraction, purification and detoxification,then adsorbed with aluminum hydroxide, filled into separate vials and lyophilized to prepare the candidate standard. The bulk and lyophilized product were tested according to the requirements in Chinese Pharmacopeia（Volume Ⅲ, 2010edition）. The candidate standard was calibrated collaboratively with WHO international standard（07/216）and Chinese national standard（002） by mouse-Vero cell antibody titration method in three laboratories. Results All the quality indexes of the candidate met the requirements for national standard. The geometric mean of 20 calibration results of potency was 289. 3 IU/ml, with 95% CI of 276. 0 ~ 303. 2 IU/ml. The calibration results with WHO international standard（07/216） and Chinese national standard（002） showed no significant difference（P 0. 05）. The result of stability test showed that the national standard for diphtheria toxoid was stable in the shelf life. Conclusion The quality indexes of prepared standard met the relevant requirements, while the potency was defined as 289. 3 IU per container. The candidate may be used as the third batch of national standard for diphtheria toxoid for potency assay.