文章摘要
林天龙.欧洲鳗免疫球蛋白单克隆抗体的制备与特性[J].水产学报,2001,25(6):
欧洲鳗免疫球蛋白单克隆抗体的制备与特性
Production and characterization of monoclonal antibodies against Anguilla anguilla IgM
投稿时间:2014-04-14  修订日期:2014-04-14
DOI:
中文关键词: 欧洲鳗鲡 单克隆抗体 免疫球蛋白 鳗鲡
英文关键词: Anguilla anguilla  monoclonal antibody  immunoglobulin
基金项目:福建省科技厅科研项目,教育部留学回国人员科研启动基金
作者单位地址
林天龙 福建省农业科学院畜牧兽医研究所 福建省农业科学院畜牧兽医研究所
摘要点击次数: 1219
全文下载次数: 1128
中文摘要:
      应用杂交瘤单克隆抗体技术制备了13个分泌抗欧洲鳗免疫球蛋白的单克隆抗体细胞株,并对这些单克隆抗体的特性进行了分析.经抗体级份和亚级份测定,其中IgM有3株,IgG1有5株,IgGa有3株,IgGb有2株;所有的单克隆抗体与欧鳗免疫球蛋白均有ELISA反应特性,抗体滴度104~107,有七株单抗具有WESTERNBLOT反应特性,能在变性条件下识别欧鳗免疫球蛋白的重链.进一步实验证实这些单抗能特异地识别欧洲鳗、日本鳗的免疫球蛋白,而与鲫、淡水白鲳、罗非鱼、胡子鲶、美洲斑点叉尾鲴血清免疫球蛋白以及水产动物常见病原菌如气单胞菌、爱德华氏菌、弧菌、柱状屈桡杆菌、沙门氏菌及大肠杆菌等无任何交叉反应.单克隆抗体9D7,对纯化的欧洲鳗免疫球蛋白的检测灵敏度为16ng.实验结果证明这些单抗具有高度特异、高度灵敏等特点,可用于鳗鲡免疫球蛋白的结构分析、免疫应答水平监测和病原诊断,具有广阔的应用前景.
英文摘要:
      By using hybridoma2monoclonal antibody technology , thirteen hybridoma cell lines which secrete monoclonal antibodies (Mab) against the IgM of European eel (Anguilla anguilla) had been established. And the characteristics of these Mabs had been described. In isotyping analysis , the results showed that among these Mabs , three of them are IgM , five of them are IgG1 , three of them are IgG2a and two of them belong to IgG2b ; all of these Mabs have the ability to bind the eel IgM by ELISA and their titer ranged from 104 to 107 , seven out of thirteen Mabs are able to recognize the heavy chain of the IgM on western blot. Further experiments proved that all of them only specifically bound to Ig of European eel and Japanese eel , and did not have any cross reaction with the Ig of Crucian carp 、Colossoma brachypomum cuvier 、Tilapia 、Clarias f uscus 、Ictalurus punctatus , and also did not react to the fish common bacterial pathogen , such as Aeromonas 、Edwardsiella 、Vibrio 、Flexibacter columnaris 、Salmonella and Escherichia coli . The measuring sensitivity of the Mab MaE9D7 to purified eel IgM was as low as 16ng. All of these results demonstrated that Mabs were highly specific and sensitive to the Ig of European eel , and had the great potential to be used for analyzing the structure of eel IgM , monitoring the immunity level of cultured eel , and pathogen diagnosis .
HTML   查看全文   下载PDF阅读器
关闭

手机扫一扫看