生产场地变更后甲型肝炎灭活疫苗(人二倍体细胞)的抗原特性

Antigenic character of inactivated hepatitis A vaccine(human diploid cells)after change of manufacturing site

目的研究生产场地变更后,原生产工艺制备的甲型肝炎灭活疫苗(人二倍体细胞)的抗原特性。方法采用双抗体夹心ELISA法检测在新场地制备的甲肝灭活疫苗病毒浓缩液和疫苗原液的甲型肝炎病毒(hepatitis A virus,HAV)抗原含量。SDS-PAGE和Western blot分析二者的蛋白组分和特异性;疫苗成品(成人剂量)稀释为不同剂量(640、320、160、80和40 EU),免疫ICR小鼠(并设铝佐剂对照组),采用竞争ELISA法检测小鼠血清中HAV抗体效价;同时称量免疫后不同时间小鼠体重。结果病毒浓缩液和疫苗原液的HAV抗原含量分别为12 800和3 200 EU/ml。疫苗原液中仅含HAV结构蛋白组分,且可与HAV抗体特异性结合。初免后28 d,640、320和160 EU剂量疫苗组小鼠血清抗体阳转率为100%,80和40 EU剂量疫苗组二免后血清抗体阳转率达100%和90%,所有免疫组小鼠血清抗体效价二免后14 d较初免28 d上升约2倍;疫苗免疫组小鼠体重及体重增幅与佐剂对照组相比,均无药物作用引起的异常变化(P>0.05)。结论在新场地采用原生产工艺制备的甲型肝炎灭活疫苗原液有单纯的抗原蛋白组分和免疫学特异性,且与前期研究结果相比,其免疫原性及安全性具有一致性。

Objective To investigate the antigenic character of inactivated hepatitis A（HA） vaccine（human diploid cells）prepared by the original procedure after change of manufacturing site. Methods The antigen contents in virus concentrate and bulk of inactivated HA vaccine produced in new site were determined by double antibody sandwich ELISA,while the protein component and specificity by SDS-PAGE and Western blot. The final product of vaccine at a dosage for adult was diluted to dosages of 640, 320, 160, 80 and 40 EU respectively and immunized to ICR mice, using aluminum adjuvant as control. The HAV antibody titer in sera of mice was determined by competitive ELISA, and the mice were weighed at various time points after immunization. Results The HAV antigen content in virus concentrate and vaccine bulk were 12 800 and 3 200 EU / ml respectively. The bulk contained only the component of HAV structural protein,which showed specific binding to HAV antibody. The serum antibody positive conversion rates of mice 28 d after immu-nization with vaccine at dosages of 640, 320 and 160 EU were 100%, while those at 80 and 40 EU were 100% and 90%respectively. The serum antibody titers of mice 14 d after the second immunization increased by about 2 folds compared with those 28 d after the first immunization. However, the bodyweights and their increases of immunized mice showed no abnormal change caused by drug as compared with those in control group（P〈0. 05）. Conclusion The bulk of inactivated HA vaccine（human diploid cells）produced in new manufacturing site showed pure protein component and immunological specificity, of which the immunogenicity and safety were consistent with those in previous study.