首批百日咳杆菌鼠源抗血清国家参考品的制备和标定

Preparation and calibration of first generation national reference for mouse antiserum to pertussis antigen

目的制备和标定百日咳杆菌鼠源抗血清国家参考品。方法按《中华人民共和国药典》2015版(三部)(简称《中国药典》)相关要求,制备候选百日咳杆菌鼠源抗血清参考品(简称候选参考品),以WHO百日咳杆菌鼠源抗血清标准品(编号97/642)为标准,组织3家单位进行协作标定,并采用热加速试验对候选参考品进行稳定性观察。结果共制备合格候选参考品300支,其外观、水分、分装精度均符合《中国药典》的相关要求;协作标定结果显示,室内和室间几何变异系数(geometric coefficient of variation,GCV)均低于20%;经统计学分析,最终确定候选参考品中含PT-Ig G、FHA-Ig G、PRN-Ig G分别为95 IU/m L、917 IU/m L、102 IU/m L,95%可信区间分别为91.7~103.7 IU/m L、900.2~944.5 IU/m L、99.4~106.1 IU/m L;每支安瓿分别含PT-Ig G 19 IU、FHA-Ig G 183 IU、PRN-Ig G21 IU;候选参考品于-20℃放置15个月及37℃放置7 d、14 d、21 d和28 d后,各种百日咳组分抗体的酶标单位值均变化不大,差异均无统计学意义(P>0.05)。结论首批百日咳杆菌鼠源抗血清国家参考品均符合《中国药典》的各项要求,且一致性、稳定性良好,可用于百日咳组分疫苗小鼠效力血清学方法的质量控制评价。

Objective To prepare and calibrate the first generation national reference for mouse antiserum to pertussis antigen. Methods According to requirement in《PHARMACOPOEIA of the People＇s Republic of China》（2015） Volume Ⅲ[simplified as《 PHARMACOPOEIA》],one batch of candidate reference for mouse antiserum to pertussis antigen（simplified as candidate） was prepared,calibrated cooperatively by 3 laboratories using WHO standard（97/642） mouse antiserum to pertussis antigen as a standard,and evaluated candidate stability by means of heat acceleration test. Results A total of300 ampoules of qualified candidate were obtained,the appearance,moisture and filling accuracy met the related requirements. Both the intra-and inter GCV（geometric coefficient of variation） of calibrated results were all less than 20%. The contents of anti-PT,anti-FHA and anti-Prn Ig G antibodies in the candidate were determined as 95 IU/m L（95%CI： 91.7-103.7 IU/m L）,917 IU/m L（95%CI： 900.2-944.5 IU/m L） and 102 IU/ml（95%CI： 99.4-106.1 IU/m L） respectively,by statistical analysis.A potency of 19 units anti-PT,183 units anti-FHA,and 21 units of anti-PRN Ig G antibodies contained in each ampoule. After stored at-20 ℃ for 15 months and at 37 ℃ for 7 d,14 d,21 d and 28 d,there were no apparent decreases observed in units of pertussis component antibodies by ELISA detection. Conclusion The first generation national reference（murine origin） met the related requirements in 《PHARMACOPOEIA》,with good consistency and stability,which may be used for the potency evaluation with serological method in mice of pertussis component vaccine.