免疫复合物型增效乙型肝炎疫苗鉴别试验方法的建立

Development of A Method for Identity Test on Immune Complex as Enhanced Hepatitis B Vaccine

目的建立免疫复合物型增效乙型肝炎疫苗鉴别试验方法。方法分别应用酸性缓冲液Gly-HCl、HAc-NaAc、Na2HPO4-CA、CA-NaCA和表面活性剂DEA-T处理免疫复合物型增效乙型肝炎疫苗,采用ELISA法检测处理后疫苗中的HBsAg和HBIG,筛选最佳缓冲液;对鉴别试验的反应条件(pH值、反应时间、反应温度)进行优化;并对鉴别试验的精密性进行验证。结果选择CA-NaCA缓冲液作为免疫复合物型增效乙型肝炎疫苗鉴别试验的样品处理缓冲液,其既具有解离抗原抗体复合物的作用,又能使铝佐剂与吸附的蛋白质解离;鉴别试验最适pH值为3.0,最佳反应温度为25℃,最佳反应时间为30 min;该鉴别试验方法精密性良好。结论 建立了免疫复合物型增效乙型肝炎疫苗鉴别试验方法,为该疫苗的检定方法提供了必要的补充。

Objective To develop a method for identity test on immune complex as enhanced hepatitis B（HB） vaccine.Methods The immune complex as enhanced HB vaccine was treated with acidic buffers Gly-HCl,HAc-NaAc,Na2HPO4-CA,CA-NaCA and surface active reagent DEA-T respectively,and determined for HBsAg and HBIG by ELISA,based on which the optimal buffer was screened.The condition for identity test,including pH value,time and temperature,was optimized.The precision of identity test was verified.Results CA-NaCA buffer was screened for treatment of samples in identity test on enhanced HB vaccine,which dissociated either antigen-antibody complex or aluminum adjuvant and the adsorbed protein.The optimal pH value,temperature and time for identity test were 3.0,25℃ and 30 min respectively.The developed method for identity test showed high precision.Conclusion A method for identity test on immune complex as enhanced HB vaccine was developed,which was a necessary supplement of determination of HB vaccine.