摘要
以基因重组技术构建工程菌株表达肠出血性大肠杆菌(EHEC)O157∶H7主要保护性抗原紧密素和志贺毒素的融合蛋白。融合蛋白采用凝胶分离电洗脱法回收纯化,用纯化的蛋白抗原免疫BALB/c小鼠,细胞融合后获得的3株杂交瘤细胞株1G2、3C6、1B10,能分别稳定分泌针对紧密素、志贺毒素Stx1和Stx2的单克隆抗体。3株单抗分别制备腹水并纯化,ELISA检测效价分别为1∶6.4×105、1∶1.2×106、1∶3200。Western-blot检测表明,3株单抗与融合蛋白发生特异性反应。应用3株单抗均可特异性检出EHECO157∶H7,而3株单抗与其他不产生紧密素和志贺毒素的大肠杆菌不反应。
The fusion proteins composed of the main protective antigens intimin and Shiga-toxin of EHEC O157 ： H7 were expressed and purified. Spleen-derived B cells from the BALB/c mouse immunized with the purified fusion protein （Eae-stxl/ 2B） were fused with SP2/0 myeloma cells and three hybridoma cell lines, named as 1G2, 3C6 and 1B10, were established by ELISA. The titre of their purified ascitic fluid were 1 ： 6. 4×10^5,1 ： 1.2×10^6 and 1 ： 3 200 in ELISA respectively. Westernblot analysis confirmed that the three McAbs only reacted specifically with fusion proteins Eae-stx1/2B or Stx1/2B respectively. The McAbs only reacted with EHEC O157 ： H7 but not with the other E. coli without intimin and Shiga-toxin.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第3期275-277,共3页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30270985)
