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目的探索氢氧化铝佐剂戊肝疫苗的免疫原性与疫苗中PO_4~(3-)浓度的关系,并分析抗体滴度与抗原在羊淋巴液中吸附率有无相关性。方法用不同浓度PO_4~(3-)处理氢氧化铝佐剂制备试验疫苗,腹腔免疫Balb/c小鼠,ELISA法定量检测免疫后不同时间血清抗体滴度,同时采用钼酸铵还原法检测试验疫苗离心沉淀中的磷含量,根据兰格缪尔方程计算抗原在含不同浓度PO_4~(3-)的佐剂表面的最大吸附量(Γm)。试验疫苗在羊淋巴液中的抗原吸附率采用ELISA法检测。结果随疫苗中PO_4~(3-)浓度增加,抗原最大吸附量(Γm)减小,抗原在羊淋巴液中的吸附率下降,免疫血清抗体滴度渐次增加,直至达到平稳状态。结论在一定范围内增加疫苗磷酸根含量可减小抗原在羊淋巴液中吸附率,降低佐剂对抗原的吸附程度,提高疫苗免疫抗体滴度。
Objective To explore the relationship between the immunogenicity of the aluminum hydroxide adjuvant hepatitis E vaccine and the concentration of PO_4 ~ (3-) in the vaccine, and to analyze whether the antibody titer is related to the antigen adsorption rate in sheep lymph fluid. Methods Balb / c mice were immunized intraperitoneally with different concentrations of PO_4 ~ (3-) -treated aluminum hydroxide adjuvant, and the serum antibody titers were measured by ELISA at different time after immunization. Meanwhile, ammonium molybdate reduction assay According to the Langmuir equation, the maximum adsorption capacity (Γm) of the antigen on the surface of the adjuvant containing different concentrations of PO_4 ~ (3-) was calculated. The antigen adsorption rate of the test vaccine in goat lymph was detected by ELISA. Results With the increasing concentration of PO_4 ~ (3-) in the vaccine, the maximum amount of antigen (Γm) decreased, the adsorption rate of antigen in goat lymph decreased, and the titer of immune serum increased gradually until it reached a steady state. Conclusion Increasing the phosphate content in the vaccine can reduce the antigen adsorption rate in goat lymph, reduce the adsorption of antigen on the adjuvant, and increase the antibody titer.