新型冠状病毒灭活疫苗抗原含量检测方法的建立

Development of method for determination the antigen content of inactivated COVID-19 vaccine

目的建立新型冠状病毒(新冠病毒)灭活疫苗抗原含量检测的双抗体夹心ELISA方法,并对该方法检测2家企业疫苗的适用性进行验证。方法使用羊抗S蛋白抗体作为包被抗体,兔抗S蛋白抗体作为显示抗体,建立了双抗体夹心ELISA方法,并对方法的准确度、精密度进行验证;分别使用该方法及企业自建方法对2家企业生产的各20批次疫苗产品进行检测,评价方法的一致性。结果成功建立了新冠病毒灭活疫苗抗原含量检测方法,该方法线性良好,R^(2)>0.99。方法验证结果显示,对于2家企业疫苗回收率均在80%~120%范围内,试验内与试验间CV均<10%。方法比对结果显示,该方法与企业方法检测结果比值对于A企业比值在0.83~1.22之间,平均为1.02;与B企业比值在0.91~1.07之间,平均为0.99;不同方法检测结果差异无统计学意义(P>0.05)。结论成功建立了新冠病毒灭活疫苗抗原含量检测方法,该方法的准确性与精密度良好,并且对国内2家企业生产的疫苗具有较好的适用性。

Objective To develop a sandwich ELISA method for determination the antigen content of inactivated COVID-19 vaccine,and verify the applicability of this method to detect vaccines from two Chinese companies.Methods A sandwich ELISA for determination of antigen content of inactivated COVID-19 vaccine was developed and the purified goat anti-S protein antibodies were used as capture antibody and rabbit anti-S protein monoclonal antibodies were used as detection antibody.The validation of the method includes accuracy and precision.The consistency between this method and the method established by other company was evaluated by testing 20 lots of vaccines from the two companies respectively.Results A sandwich ELISA for determination the antigen content of inactivated COVID-19 vaccine was established which has good linearity with R^(2)>0.99.The recovery rates for vaccines from two Chinese companies were all within the range of 80%-120%.The CVs of intra-and inter-assays were both less than 10%.For 20 lots of vaccines from A company,the ratios of results tested by two different methods were ranged from 0.88 to 1.20,with an average of 1.02.For 20 lots of vaccines from B company,the ratios were ranged from 0.91 to 1.07,with an average of 0.99.There was no statistical difference in the results from different methods(P>0.05).Conclusion A sandwich ELISA method for determination of antigen content of inactivated COVID-19 vaccine was successfully established,and this method has good accuracy and precision,and is applicable to determine the antigen content of vaccines from two Chinese companies.