氢氧化铝佐剂对重组大肠杆菌表达的戊型肝炎239抗原的吸附研究  

Research on the adsorption of HEV239 expressed on recombinant Escherichia. Coli by aluminum hydroxide adjuvant

作　　者：田俊楠 叶祥忠 张娜 张明祥 鲜阳凌 李益民 张景海[2] 

机构地区：[1]北京万泰生物药业股份有限公司,102206 [2]沈阳药科大学生化教研室,110016

出　　处：《免疫学杂志》2010年第6期465-468,共4页Immunological Journal

基　　金：国家高技术研究与发展项目863计划(2005AA2Z3H20)

摘　　要：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。We explored the adsorption force by which hepatitis E antigen expressed in recombinant Escherichia. Coli （HEV239） adsorbed on aluminum hydroxide adjuvant, and its application in preparation of HEV vaccine. By using Langmuir adsorption equation, we calculated the adsorption capacity （Γm） of HEV239 upon aluminum hydroxide adjuvant in the presence of different magnitudes of sodium chloride （NaCl） or ethylene glycol （EG）. Then we estimated the kind of adsorption force by the tendency of Γm when concen-tration of NaCl or EG changed. Experimental vaccines were prepared which containing polyhydroxy-compound and different quantities of phosphate, and then plotted Γms line graph as phosphate changed. After the experimental vaccines were mixed with sheep lymph fluid for 12 h, the adsorption percentage of antigen was detected quantitatively by ELISA. We found that Γm of HEV239 decreased as ionic strength （concentration of NaCl） or amount of hydrophobic material （volume of EG） increased, so do it when phosphate in vaccines increased. The adsorption percentage of HEV239 decreased until reached a stable state when phosphate of experimental vaccines increased. Our results indicate that both electrostatic attractive force and hydrophobic force contribute to the adsorption of HEV239 on aluminum hydroxide adjuvant. Based on this mechanism, adding phosphate and polyhydroxy compound to experimental vaccines can down-regulate Γm in formulation and adsorption capacity in sheep lymph fluid.

关 键 词：氢氧化铝佐剂 吸附力 吸附率 羊淋巴液 

分 类 号：R392[医药卫生—免疫学]