| 鼠疫耶尔森氏菌Pla蛋白单克隆抗体的制备与鉴定 |
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| 引用本文： | 杜春红,王鹏,张建中,唐雪,宋志忠.鼠疫耶尔森氏菌Pla蛋白单克隆抗体的制备与鉴定[J].细胞与分子免疫学杂志,2011,27(9):985-988. |
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| 作者姓名： | 杜春红 王鹏 张建中 唐雪 宋志忠 |
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| 作者单位： | 1. 云南省地方病防治所,云南大理,671000
2. 中国疾病预防控制中心传染病预防控制所,北京,102206 |
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| 基金项目： | 云南省科技攻关项目(2007CA010) |
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| 摘 要： | 目的:利用原核表达的鼠疫耶尔森氏菌Pla蛋白(rPla),通过杂交瘤细胞技术制备单克隆抗体(mAb),为相关研究工作奠定基础.方法:大肠杆菌表达的Pla蛋白,包涵体经尿素反复洗涤纯化后免疫BALB/c小鼠,取血清抗体效价高的小鼠脾细胞与Sp2/0骨髓瘤细胞进行细胞融合,以表达Pla、表达GST、天然提取Pla 3种蛋白为抗原物,采用间接ELISA法筛选阳性杂交瘤细胞,并结合Western blot对所获取mAb的特异性进行鉴定.结果:经间接ELISA筛选,获得3株能稳定分泌抗天然Pla蛋白mAb的杂交瘤细胞株,命名为15B8、14H4、19A4,亚类测定分别为IgG2a和IgG1,轻链均为κ链;腹水经间接ELISA法检测效价可达1:106:Western blot实验证实该3株mAb能特异性识别天然Pla蛋白.结论:成功获得了抗鼠疫耶尔森氏菌天然Pla抗原的特异性mAb,为进一步研究Pla蛋白及研发诊断试剂奠定了基础.
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| 关 键 词： | 鼠疫耶尔森氏菌 Pla蛋白 单克隆抗体 |
Preparation and identification of monoclonal antibodies against Pla protein of Yersinia pestis |
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| DU Chun-Hong,Wang Peng,Zhang Jian-Zhong,Tang Xue,Song Zhi-Zhong.Preparation and identification of monoclonal antibodies against Pla protein of Yersinia pestis[J].Journal of Cellular and Molecular Immunology,2011,27(9):985-988. |
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| Authors: | DU Chun-Hong Wang Peng Zhang Jian-Zhong Tang Xue Song Zhi-Zhong |
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| Affiliation: | DU Chun-hong1,WANG Peng1,ZHANG Jian-zhong2,TANG Xue1,SONG Zhi-zhong1 1Yunnan Institute for Endemic Diseases Control and Prevention,Dali 671000,2National Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and Prevention,Beijing 102206,China |
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| Abstract: | AIM: To prepare the monoclonal antibody of Pla with recombinant Pla(rPla) by hybridoma cell technology,which will lay the foundation for related research work.METHODS: Purified rPla was collected by washing repeatedly with urea,and BALB/c mice were immunized by them.Hybridoma cells were achieved by Sp2/0 cell fusion with mouse spleen cells from successfully immunized mice.Monoclonal antibody was screened by indirect ELISA and Western blots with rPla,natural crude Pla and GST respectively.RESULTS: Three stra... |
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| Keywords: | Yersinia pestis Pla protein monoclonal antibody |
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