欧洲鳗免疫球蛋白单克隆抗体的制备与特性

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，IgG2a有3株，IgG2b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10^4-10^7，有七株单抗具有WESTERN BLOT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Hui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。

Production and characterization of monoclonal antibodies against Anguilla anguilla IgM

By using hybridoma monoclonal 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 IgG 1, three of them are IgG 2a and two of them belong to IgG 2b ; all of these Mabs have the ability to bind the eel IgM by ELISA and their titer ranged from 10 4 to 10 7, 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 fuscus?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.