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目的:探索氢氧化铝佐剂对重组大肠杆菌表达的戊型肝炎239抗原(HEV239)的吸附力种类以及在戊肝疫苗制备研究中的应用。方法:用兰格缪尔吸附方程计算在不同浓度氯化钠(NaCl)或乙二烯乙二醇(EG)条件下,HEV239在氢氧化铝佐剂表面的最大吸附量(Γm),根据Γm随NaCl或EG浓度变化趋势判断吸附作用力的种类。配制添加多羟基化合物且含不同浓度磷酸盐的试验疫苗,计算Γm并绘制其随磷浓度变化的曲线。ELISA法定量检测并计算含不同浓度磷的试验疫苗与羊淋巴液作用12h后抗原的吸附率。结果:HEV239的Γm随离子强度(NaCl浓度)或疏水物质浓度(EG体积)增加而降低,也随磷酸盐浓度增加而降低。疫苗在羊淋巴液中的抗原吸附率随磷酸盐浓度增加而降低,最终达到稳定水平。结论:HEV239在氢氧化铝佐剂表面的吸附受到静电引力和疏水作用力的双重影响。利用此机理向疫苗中添加磷酸盐和多羟基化合物可降低疫苗抗原在制剂中的吸附量和在羊淋巴液中的吸附率。
Objective: To explore the adsorption of aluminum hydroxide adjuvant on recombinant hepatitis E 239 antigen (HEV239) expressed in E. coli and its application in the preparation of hepatitis E vaccine. Methods: The maximum adsorption capacity (Γm) of HEV239 on the surface of aluminum hydroxide adjuvant was calculated with Langmuir adsorption equation under different concentrations of sodium chloride (NaCl) or ethylene glycol (EG). According to Γm With the NaCl or EG concentration trend to determine the type of adsorption force. Formulations of test vaccines supplemented with polyhydroxyl compounds and containing varying concentrations of phosphate were calculated and plotted as a function of phosphorus concentration. The ELISA method was used to quantitatively detect and calculate the adsorption rate of the antigen after 12 hours of experiment vaccine containing different concentrations of phosphorus and sheep lymph fluid. Results: The Γm of HEV239 decreased with increasing ionic strength (NaCl concentration) or hydrophobic material concentration (EG volume), also decreasing with increasing phosphate concentration. The antigen adsorption rate of the vaccine in goat lymph decreased with the increase of phosphate concentration and eventually reached a stable level. Conclusion: The adsorption of HEV239 on the surface of aluminum hydroxide adjuvant is affected by both electrostatic attraction and hydrophobic interaction. Using this mechanism to add phosphates and polyols to the vaccine reduces the amount of vaccine antigen adsorbed in the formulation and the rate of adsorption in the sheep lymph fluid.