| 基因免疫制备牛分枝杆菌MPB64抗原单克隆抗体 |
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| 引用本文： | 刘建东,刘思国,王春来,宫强,王勇,迟磊,赵昆. 基因免疫制备牛分枝杆菌MPB64抗原单克隆抗体[J]. 中国预防兽医学报, 2007, 29(11): 870-873 |
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| 作者姓名： | 刘建东 刘思国 王春来 宫强 王勇 迟磊 赵昆 |
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| 作者单位： | 1. 中国农业科学院哈尔滨兽医研究所,国家兽医生物技术重点实验室/动物细菌病研究室,黑龙江,哈尔滨,150001
2. 东北农业大学,黑龙江,哈尔滨,150001 |
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| 摘 要： | 为了建立针对牛分枝杆菌分泌抗原MPB64的检测手段,制备该抗原的单克隆抗体,构建真核表达载体pcDNA-mpb64,以原核表达并纯化的融合蛋白H-MPB64包被ELISA板,建立单克隆抗体检测方法。利用基因免疫方法免疫BALB/c小鼠,通过杂交瘤技术筛选出针对牛分枝杆菌分泌蛋白MPB64的单克隆抗体一株,并制备了腹水,经鉴定腹水效价达到1∶10240,采用硫酸铵沉淀法纯化腹水,纯化抗体效价为1∶8000,为今后研制分枝杆菌鉴定技术奠定基础。
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| 关 键 词： | 牛分枝杆菌MPB64蛋白 基因免疫 单克隆抗体 |
| 文章编号： | 1008-0589（2007）11-0870-04 |
| 修稿时间： | 2007-03-15 |
Preparation of monoclonal antibodies against the MPB64 antigen of Mycobacterium bovis by gene immunization |
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| LIU Jian-dong,LIU Si-guo,WANG Chun-lai,GONG Qiang,WANG Yong,CHI Lei,ZHAO Kun. Preparation of monoclonal antibodies against the MPB64 antigen of Mycobacterium bovis by gene immunization[J]. Chinese Journal of Preventive Veterinary Medicine, 2007, 29(11): 870-873 |
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| Authors: | LIU Jian-dong LIU Si-guo WANG Chun-lai GONG Qiang WANG Yong CHI Lei ZHAO Kun |
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| Affiliation: | 1. Division of Bacterium Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China; 2. Northeast Agricultural University, Harbin 150001, China |
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| Abstract: | To establish a new detection method for MPB64 antigen of M.bovis,a mammalian expression vector pcDNA- mpb64 was constructed and used to immunize the BALB/c mice.A prokaryotic system expressing H-MPB64 protein as detective antigen was used for testing monoclonal antibodies against the MPB64 antigen of M.bovis.One hybridoma cell line secreting monoclonal antibodies was established.The McAb reacted with H-MPB64 and PPD in ELISA experiment,but not in western blot. |
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| Keywords: | Mycobacterium bovis gene immunization McAb |
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