目的采用双抗体夹心ELISA法检测鼠疫组分疫苗中F1和rV的抗原含量。方法利用F1、rV抗原单克隆抗体,对鼠疫组分疫苗原液进行抗原鉴别试验;分别应用F1、rV抗原双抗体夹心ELISA法对疫苗原液进行抗原定量检测;验证A(l OH)3佐剂吸附抗原的完全性;将疫苗成品于4℃放置18个月,分别于1和18个月时取样,用建立的双抗体夹心ELISA法检测F1和rV抗原含量,分析抗原的稳定性;对3批鼠疫菌rV抗原原液3步纯化工艺进行验证。结果鼠疫组分疫苗原液中的F1和rV抗原具有良好的反应原性;用建立的双抗体夹心ELISA法检测4批F1和rV抗原原液中F1和rV抗原的含量与Lowry法检测结果的误差均在±20%以内;4批鼠疫组分疫苗离心后的上清液中均未检测到F1和rV抗原,表明A(lOH)3佐剂吸附抗原完全;4℃保存18个月,疫苗中的F1和rV抗原含量稳定;3批鼠疫菌rV抗原原液经阴离子交换层析、疏水层析、凝胶过滤层析3步纯化,rV抗原在纯度增加的同时,抗原含量也增加。结论已建立的F1、rV抗原双抗体夹心ELISA法可用于鼠疫组分疫苗中F1和rV抗原的定量检测。 Objective To detect the antigenic content of F1 and rV in the plague vaccine by double antibody sandwich ELISA. Methods Monoclonal antibodies against F1 and rV were used to test the antigenicity of the vaccine against the plague components. Antigen was detected by double antibody sandwich ELISA with F1 and rV antigens respectively. The adsorption of A (lOH) 3 adjuvant The completeness of the antigen was tested. The final vaccine product was placed at 4 ℃ for 18 months and sampled at 1 and 18 months respectively. The antibody levels of F1 and rV were detected by double antibody sandwich ELISA. The stability of the antigen was analyzed. Yersinia pestis rV antigen stock solution 3-step purification process to verify. Results The F1 and rV antigens of the plague component vaccine liquid had good reactionogenicity. The double antibody sandwich ELISA was used to detect the difference of the F1 and rV antigens between the four batches of F1 and rV antigens and the Lowry test Within ± 20%; F1 and rV antigens were not detected in the supernatant of the four batches of the plague component vaccine after centrifugation, indicating that the antigen adsorbed by the A (lOH) 3 adjuvant was completely intact; preserved at 4 ° C for 18 months, The contents of F1 and rV antigens were stable. Three batches of Y. pestis rV antigen solution were purified by anion exchange chromatography, hydrophobic chromatography and gel filtration chromatography. The antigen content of rV antigen was also increased with the increase of purity. Conclusion The established sandwich ELISA of F1 and rV antigens can be used for the quantitative detection of F1 and rV antigens in the plague vaccine.