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当前位置: 首页 > 文章 > 动物源性人兽共患细菌病防控生物新制剂的研究Ⅰ.分泌抗大肠杆菌O157∶H7单克隆抗体杂交瘤细胞株的建立 江苏农业学报 2009,25 (2) 73-77
Position: Home > Articles > Novel Biological Products for Prevention and Control of Bacterial Zoonoses Ⅰ. Establishment of the Hybridoma Producing Monoclonal Antibodies against Enterohemorrhagic E. coli O157∶H7 Jiangsu Journal of Agricultural Sciences 2009,25 (2) 73-77

动物源性人兽共患细菌病防控生物新制剂的研究Ⅰ.分泌抗大肠杆菌O157∶H7单克隆抗体杂交瘤细胞株的建立

作  者：
夏兴霞;刘洁;王永山;诸玉梅;欧阳伟;何孔旺;张雪寒
单  位：
江苏省农业科学院兽医研究所
关键词：
肠出血性大肠杆菌O157∶H7;单克隆抗体(mAb);生物制剂;人兽共患病
摘  要：
将E.coliO157∶H7(EHEC-O157∶H7)用甲醛灭活、超声破碎,免疫BALB/c小鼠,制备免疫脾细胞;与SP2/0骨髓瘤细胞融合后,获得7株分泌抗E.coliO157∶H7单克隆抗体(mAbs)的杂交瘤细胞株,分别命名为F5、B8、E7、G9、G11、F1和C3,Ig亚类分别是IgG2aκ、IgG2aκ、IgG1κ、IgMκ、IgMλ、IgMκ和IgG2aκ。用间接ELISA检测,E7和F1细胞培养上清液的效价为1.6×103,腹水效价均为1×1011;C3、F5、B8、G9和G11细胞培养上清液的效价为1×10~2×102,腹水效价均为1×103。相加ELISA结果显示,F1、G9和G11 3株mAbs对应相同的抗原表位,而其它4株对应不同的抗原表位。用间接ELISA分析特异性,7株mAbs与某些其他血清型的大肠杆菌有交叉反应。Western blotting表明,C3、E7和F1 3株mAbs在蛋白分子量28×103处有特异性条带,而其它4株mAbs则未显示蛋白带。
译  名：
Novel Biological Products for Prevention and Control of Bacterial Zoonoses Ⅰ. Establishment of the Hybridoma Producing Monoclonal Antibodies against Enterohemorrhagic E. coli O157∶H7
作  者：
XIA Xing-xia1,LIU jie1,2,WANG Yong-shan1,ZHU Yu-mei1,OUYANG Wei1,2,HE Kong-wang1,ZHANG Xue-han1(1.Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Key Laboratory of Animal Diseases Diagonostic and Immunology,Ministry of Agriculture,National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China;2.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)
关键词：
enterohemorrhagic E.coli O157∶H7(EHEC-O157∶H7);monoclonal antibody(mAb);biological product;zoonoses
摘  要：
Seven hybridomas secreting monoclonal antibodies(mAbs) against enterohemorrhagic E.coli O157∶H7(EHEC-O157∶H7)were established by fusing SP2/0 with spleen cells from BALB/c mice immunized with formaldehyde-killed and ultrasonically-broken E.coli O157∶H7,naming E7,F1,C3,F5,B8,G9 and G11.The Ig isotypes of mAbs were IgG2aκ,IgG2aκ,IgG1κ,IgMκ,IgMλ,IgMκ and IgG2aκ,respectively.The ELISA antibody titers of E7 and F1 were 1.6×103 and 1×1011in supernatant and ascitic fluid respectively,the others were 1×10-2×102 in supernatant,and 1×103 in the ascitic fluids.Additivity ELISA indicated that F1,G9 and G11 were corresponding to the same antigen epitope,but the other 4 mAbs were different epitopes.The 7 mAbs could react with some other kinds of bacteria in the specific analysis by indirect ELISA.In the western-blotting,mAbs C3,E7 and F1 displayed thestained protein band of approximate 28×103,but the other 4 mAbs didn't show any stained protein band.

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