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当前位置: 首页 > 文章 > 肠出血性大肠杆菌O157:H7紧密素单克隆抗体的制备及鉴定 畜牧与兽医 2014,46 (3) 37-40
Position: Home > Articles > Preparation and identification of the monoclonal antibody against intimin of Escherichia coli O157: H7 Animal Husbandry & Veterinary Medicine 2014,46 (3) 37-40

肠出血性大肠杆菌O157:H7紧密素单克隆抗体的制备及鉴定

作  者：
李盟;张雪寒;何孔旺;黄克和
单  位：
江苏省农科院兽医所人兽共患病实验室;南京农业大学动物医学院
关键词：
肠出血性大肠杆菌O157:H7;紧密素;单克隆抗体
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O157:H7紧密素的融合蛋白。用纯化的紧密素蛋白免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,对杂交瘤细胞及时筛选,阳性孔经4次有限稀释法克隆,成功获得2株能稳定传代并分泌抗紧密素单克隆抗体(McAb)的杂交瘤细胞株。2株单抗分别制备腹水,ELISA检测效价分别为5.2×104,2.5×104。Western blot检测表明,2株单抗与融合蛋白发生特异性反应;ELISA检测表明,2株单抗可特异性检出大肠杆菌O157:H7。本研究为建立大肠杆菌O157检测方法提供了物质基础。
译  名：
Preparation and identification of the monoclonal antibody against intimin of Escherichia coli O157: H7
作  者：
LI Meng;ZHANG Xuehan;HE Kongwang;HUANG Kehe;College of Veterinary Medicine,Nanjing Agricultural University;Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences;
关键词：
E.coli O157: H7;;intimin;;monoclonal antibody
摘  要：
The recombinant intimin protein of Escherichia coli O157: H7 was obtained by genetic engineering technology. Spleen cells of mice immunized with the purified intimin were fused with SP2 /0 myeloma cells to generate hybridomas. Hybridomas were screened timely and positive wells were cloned four times by limiting dilution. Two stable hybridoma cell lines secreting the antibody to intimin were obtained successfully. The titers of the ascetic were 5. 2 × 104and 2. 5 × 104,respectively. Western-blot analysis showed that two monoclonal antibodies( mAbs) could react with the recombinant protein. ELISA demonstrated that the mAbs were specifically against E. coli O157: H7. The study provides a basis for development of detection method for E. coli O157.

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