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| 抗OmpW单克隆抗体的制备与特性鉴定 | |
| 引用本文： | 马振宁,郑磊,林海关,郑允权,唐凤翔,石贤爱,郭养浩.抗OmpW单克隆抗体的制备与特性鉴定[J].细胞与分子免疫学杂志,2011,27(9):982-984. |
| 作者姓名： | 马振宁 郑磊 林海关 郑允权 唐凤翔 石贤爱 郭养浩 |
| 作者单位： | 1. 福州大学药物生物技术与工程研究所 2. 福建省医疗器械与医药技术重点实验室,福建福州,350002 3. 福州大学药物生物技术与工程研究所;福建省医疗器械与医药技术重点实验室,福建福州350002 |
| 基金项目： | 福建省科技厅重点项目(2009N401010844);福建省科技厅重点项目(2010N0014); 福建省教育厅科技项目(JA10024) |
| 摘 要： | 目的:制备抗副溶血弧菌OmpW的单克隆抗体(mAb),并鉴定其特性.方法:利用生物信息学方法分析多种病原弧菌的OmpW氨基酸序列,原核表达并纯化制备r-OmpW.以r-OmpW作为免疫抗原制备鼠mAb,以五种病原弧菌为筛选抗原.采用Western blot、流式细胞术(FCM)、间接ELISA法鉴定mAb的特性.结果:生物信息学分析表明OmpW是弧菌内高度保守的外膜蛋白.成功筛选到3株杂交瘤细胞株,其中,杂交瘤细胞株S5C10分泌的mAb的Ig亚类(型)为IgG3,腹水mAb效价达4.6×104.该mAb同时识别副溶血弧菌、溶藻弧菌、哈维氏弧菌、鳗弧菌和创伤弧菌,而与荧光假单胞菌、温和气单胞菌、嗜水气单胞菌、豚鼠气单胞菌及大肠杆菌无明显的交叉反应.结论:制备的mAb可特异性识别上述5种病原弧菌,为进一步建立广谱弧菌的免疫学检测方法提供了工具. |
| 关 键 词： | OmpW 弧菌 原核表达 单克隆抗体 流式细胞术 |
Preparation and identification of OmpW monoclonal antibodies |
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| Ma Zhen-Ning,Zheng Lei,Lin Hai-Mei,Zheng Yun-Quan,Tang Feng-Xiang,Shi Xian-Ai,Guo Yang-Hao.Preparation and identification of OmpW monoclonal antibodies[J].Journal of Cellular and Molecular Immunology,2011,27(9):982-984. | |
| Authors: | Ma Zhen-Ning Zheng Lei Lin Hai-Mei Zheng Yun-Quan Tang Feng-Xiang Shi Xian-Ai Guo Yang-Hao |
| Institution: | MA Zhen-ning1,ZHENG Lei1,LIN Hai-mei1,ZHENG Yun-quan2,TANG Feng-xiang2,SHI Xian-ai2,GUO Yang-hao1,21Institute of Pharmaceutical Biotechnology and Bioengineering,Fuzhou University,2Fujian Key Lab of Medical Instrumentation and Pharmaceutical Technology,Fuzhou 350002,China |
| Abstract: | AIM: To prepare and characterize the mouse monoclonal antibodies against Vibrio parahaemolyticus OmpW.METHODS: The OmpW amino acid sequence from three diseased Vibrio was analyzed by Bioinformatics.Mice were immunized with r-OmpW which was highly expressed and purified in E.coli.Five Vibrio(Va,Vp,Vh,Vv,Van) were chosen as antigen for mAb selection.The characters of the anti-OmpW monoclonal antibodies were studied by Western blot,Flow Cytometry,indirect immunofluorescence.RESULTS: OmpW was testified a highly... |
| Keywords: | OmpW Vibrio prokaryotic expression monoclonal antibody flow cytometry |
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