酶联免疫法测定重组乙型肝炎疫苗中庆大霉素残留量的相关影响因素分析

Analysis of influencing factors of determination of residual gentamicin in recombinant hepatitis B vaccine by ELISA

目的分析酶联免疫法测定重组乙型肝炎疫苗中庆大霉素残留量的相关影响因素。方法使用庆大霉素酶联免疫试剂盒对3种表达系统的重组乙型肝炎疫苗（酿酒酵母、汉逊酵母、CHO细胞）成品和原液进行庆大霉素回收率测定，同时对不同来源庆大霉素标准品绘制的标准曲线进行对比，对铝佐剂和标准品对庆大霉素残留量检测的影响进行分析。结果含铝佐剂的乙型肝炎疫苗成品中庆大霉素回收率差异较大，疫苗原液中测定庆大霉素的回收率较稳定，在85．1％-101．5％之间，波动性较小；试剂盒自带标准品与庆大霉素国家标准品绘制的标准曲线存在差异。结论不同制备工艺的铝佐剂会影响乙型肝炎疫苗中庆大霉素的回收率，在原液中测定庆大霉素残留量稳定性良好，标准品来源及批次会影响庆大霉素残留量的测定。

Objective To investigate the influencing factors of determination of residual gentamicin in recombinant hepatitis B （HB） vaccine by ELISA. Methods The bulks and final products of recombinant HB vaccine expressed in three systems （Saccharomyces cerevisiae, Hansenula anomala and CHO cells） were determined for recovery rates of gentamicin by ELISA kit. The standard curves of gentamicin from various origins were compared, based on which the influences of albumin adjuvant and standard on determination of residual gentamicin were analyzed. Results The recovery rates of gentamicin were varied in final product of aluminum adjuvant-containing HB vaccine, while were stable as 85. 1% - 101. 5% in bulk. However, the standard curve plotted with the standard gentamicin in the kit was different from that with national standard gentamicin. Conclusion Aluminum adjuvant manufactured by various procedures influenced the recovery rate of gentamicin in HB vaccine. The residual gentamicin content in bulk of vaccine was stable. However, the determination results by using standards of various origins and batches were different.