Asia I型口蹄疫疫苗效力检验替代方法的初步研究

使用三株针对AsiaⅠ型口蹄疫146S抗原的杂交瘤细胞株(1#G3C2、2#G6B9、10#C2G1)大量生产腹水单克隆抗体,粗提纯化并按一定的比例进行混合。初步建立混合腹水单克隆抗体介导的间接夹心ELISA方法,使用该方法测定AsiaⅠ型病毒灭活抗原、O型病毒灭活抗原、正常BHK21细胞培养液,结果表明该方法特异性好,所测定OD值和AsiaⅠ型抗原稀释倍数呈明显线性关系。用该方法对11批AsiaⅠ型、O型单价或双价成品疫苗破乳抗原进行测定,结果表明不同批次不同企业生产的疫苗完整病毒颗粒抗原含量有差异,进一步完善有望用于AsiaⅠ型口蹄疫成品疫苗中有效颗粒抗原含量的测定,从而发展成一种疫苗效力检验替代方法。

Study on an Alternative Approach for Assessment of Asia I Foot-and-Mouth Disease Vaccine Potency

Three hybridoma cell line(1#G3C2、2#G6B9、10#C2G1) secreting stably monoclonal antibody against 146S antigen of Asia I FMDV were prepared.These monoclonal antibodies were purified and mixed by a certain proportion.An IS-ELISA based on the mixture of the three different monoclonal antibodies was developed and applied to the testing of the inactivated Asia I FMDV antigens,inactivated O FMDV antigens and BHK21 cell supernatant.The experiment result demonstrated that the assay was highly specific and there was a clear linear relationship between the determined OD value and Asia I FMDV antigen dilutions.The assay was used to test total 11 batches of demulsified antigens of FMD vaccines,including Asia I,O and bivalent(O-Asia I) and the data indicated that the whole virus particle contents i.e.146S antigen contents were different among different commercial vaccines.It conld be expected to test the effective antigen yields in commercial Asia I FMD vaccines and become an alternative means for assessment of vaccine potency.