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Position: Home > Articles > Development and Identification of the Monoclonal Antibodies against the Main Protective Antigens of EHEC O157∶H7 Chinese Journal of Veterinary Science 2006 (3) 275-277

肠出血性大肠杆菌O157∶H7主要保护性抗原单克隆抗体的研制及特性分析

作  者:
祝令伟;冯书章;刘军;陈萍
单  位:
军事医学科学院军事兽医研究所
关键词:
肠出血性大肠杆菌O157∶H7;单克隆抗体;紧密素;志贺毒素
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。
译  名:
Development and Identification of the Monoclonal Antibodies against the Main Protective Antigens of EHEC O157∶H7
作  者:
ZHU Ling-wei, FENH Shu-zhang, LIU Jun, CHEN Ping (The Military Veterinary Institute, Academy of Military Medicine of PLA, Changchun 130062, China)
关键词:
EHEC O157∶H7;monoclonal antibody;intimin;Shiga-toxin
摘  要:
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-stx1/2B) 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∶6.4×105,1∶1.2×106 and 1∶3 200 in ELISA respectively. Western-blot 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.

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