【摘要】 :(1)依据 p H6抗原的生化特征及表达条件 ,对不同温度 (37℃和 2 8℃ )及不同酸碱度 (p H6和 p H7)条件下培养细菌的提取物进行了电泳图型对比 ;(2 )依照上述电泳图型 ,并结合该抗原的分子量 ,确定了 p H6抗原的电泳迁移位置 ;(3)采用 KCNS洗脱、(NH4) 2 SO4盐析、制备电泳的方法提取和纯化了鼠疫耶尔森氏菌 (Yersinia pestis) p H6抗原 ,SDS-PAGE分析表明 ,提取物均一 ,表征分子量符合 ,回收率较高 ;(4)使用纯化抗原免疫普通小鼠 5只、BAL B/ c小鼠 3只 ,免疫效果良好 ,利用获得的免疫血清 ,建立了 p H6抗原多克隆抗体的 EL ISA检测技术 ,应用于融合细胞中阳性克隆的筛检及实验动物的免疫效果评价 ,应用结果证实 ,检测敏感、有效 ;(5 )利用细胞融合技术获得杂交瘤 17株 ,对其中的 4株进行了 3次再克隆并经半年反复冷冻、复苏及上清检测 ,证实瘤株分泌抗体稳定 ;(6 )对所获单抗进行了 Western blotting(蛋白印迹试验 )鉴定 ,结果显示抗体特异。更多还原

【Abstract】 According to the biochemistry characteristic and expressing condition of pH6 antigen, electrophoresis contrast was made under different temperature(37℃ and 28℃) and different pH(pH6 and pH7). (2) According to electrophoresis photograph and the molecular weight of pH6 antigen, the position in electrophoresis photograph was labled. (3) The pH6 antigen of Yersinia pestis was purified during KCNS washing, SAS precipitation and electrophoresis preparation. The SDS-PAGE analysis indicated that the collecting be homogeneous, the appearance molecular weight tally with the facts and the rate of recovery be higher. (4) Five normal mice and three BALB/c mice were injected by the purified pH6 antigen. The immune result was satisfactory. The ELISA technique was established with immune serum. It proved that the technique be sensitive and qualified when used in selecting positive clone cell and examing immune effect. (5) There were 17 strains of hybridization cell gotten. Four strains of stable hybridizaition cell were selected from 17 strains of hybridization cell. (6) The obtained McAb was exceptionally proved by western blotting.更多还原