| 牛型布氏杆菌苹果酸脱氢酶单克隆抗体的制备及鉴定 |
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| 引用本文： | 童永亮,韩先干,孙晓庆,丁铲,胡青海,丁家波,王豪举,于圣青. 牛型布氏杆菌苹果酸脱氢酶单克隆抗体的制备及鉴定[J]. 中国兽医科学, 2012, 0(6): 596-600 |
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| 作者姓名： | 童永亮 韩先干 孙晓庆 丁铲 胡青海 丁家波 王豪举 于圣青 |
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| 作者单位： | 西南大学动物科技学院;中国农业科学院上海兽医研究所;中国兽医药品监察所 |
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| 基金项目： | 国家重点基础研究发展计划(973)项目(2010CB530202);中央级科研院所公益性研究专项(2012JB05) |
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| 摘 要： | 为制备牛型布氏杆菌(Brucella abortus)苹果酸脱氢酶(MDH)单克隆抗体,将牛型布氏杆菌A19菌株的MDH基因进行克隆和表达,表达产物被纯化后作为免疫原,按常规方法免疫BALB/c小鼠。取免疫鼠脾细胞与SP2/0骨髓瘤细胞融合,经亚克隆和间接ELISA筛选,最终获得3株分泌牛型布氏杆菌MDH蛋白单克隆抗体的杂交瘤细胞,分别被命名为C2、D5和F4。ELISA检测结果表明,3株杂交瘤细胞培养上清的抗体效价为1∶1 600～1∶3 200,小鼠腹水效价为1∶12 800～1∶25 600;Western-blot检测结果表明,3株单克隆抗体均能与牛型布氏杆菌MDH重组蛋白发生特异性反应。结果表明,本研究成功制备了牛型布氏杆菌MDH蛋白单克隆抗体,为基于MDH的布氏杆菌检测方法的建立奠定了基础。
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| 关 键 词： | 牛型布氏杆菌 苹果酸脱氢酶 单克隆抗体 |
Preparation and characterization of monoclonal antibodies against Brucella abortus MDH recombinant protein |
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| TONG Yong-liang,HAN Xian-gan,SUN Xiao-qing,DING Chan,HU Qing-hai,DING Jia-bo,WANG Hao-ju,YU Sheng-qing. Preparation and characterization of monoclonal antibodies against Brucella abortus MDH recombinant protein[J]. Chinese Veterinary Science, 2012, 0(6): 596-600 |
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| Authors: | TONG Yong-liang HAN Xian-gan SUN Xiao-qing DING Chan HU Qing-hai DING Jia-bo WANG Hao-ju YU Sheng-qing |
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| Affiliation: | 1.College of Animal Science and Technology,Southwest University,Chongqing 400716,China;2.Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,China; 3.China Institute of Veterinary Drug Control,Beijing 100081,China) |
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| Abstract: | The malate dehydrogenase(MDH) of Brucella abortus A19 was expressed,purified and used for immunization to BALB/c mice.The spleen cells of the immunized BALB/c mice were collected and fused with SP2/0 cells,after three sub-clones.Three hybridoma cell strains C2,D5 and F4,which secret specific monoclonal antibodies(MAbs) against MDH,were obtained by ELISA.The antibody titers in the cell culture supernatant and ascites were from 1∶1 600 to 1∶3 200 and from 1∶12 800 to 1∶25 600,respectively.Western-blot analysis showed that three MAbs reacted with expressed brucella MDH protein specifically,which provided basis for further study of MDH. |
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| Keywords: | Brucella abortus malate dehydrogenase monoclonal antibody |
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