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Analysis of immune-reactive components of ayu serum using monoclonal antibody

使用单克隆抗体分析香鱼血清的免疫反应成分

基本信息

批准号：
62560195
负责人：
KAWAI Kenji
金额：
$ 1.34万
依托单位：
Kochi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560195/
关键词：
Monoclonal Antibody Immune-reactive Component Ayu 血清成分免疫ハイブリドーマ

项目摘要

The subject of the present study was to make monoclonal antibodies against serum components especially main immune reactive components of ayu, Plecoglossus altivelis, to detect and quantitatively determine those components.Trials to make monoclonal antibodies were done repeatedly by screening and cloning hybridomas of established mouse myeloma cells and lymphocytes obtained from the spleen of mice immunized with ayu serum by means of immune blotting and enzyme-linked immunosorbent assay (ELISA) with polystyrene beads sensitized with ayu serum using anti-mouse IgG labeled with peroxydase. To make anti-ayu antibody monoclonal antibody, ELISA using the serum of ayu immunized with Vibrio anguillarum and polystyrene beads sensitized with V.anguillarum lipopolysaccharide. Detection was done on the serum and homogenates of the kidney, spleen and liver of immunized and unimmunized ayu. Detection was dine also on eggs and fry of ayu obtained from a hatchery.Totally twelve monoclonal antibodies ("A"-"L") were obtained. Monoclonal antibody "A" was considered to be that against ayu antibody. Antigens "a"-'1", which corresponded to monoclonal antibodies "A"-"L", were detected in the serum and the homogenates of the kidney, spleen and liver and the fries but only "b"-"e" and "1" were detected in the eggs. Concentration of the "a" was higher in the immunized ayu than that in the unimmumized ayu, but the concentration of other antigens did not increasae by immunization of ayu.

本研究的目的是制备针对阿玉主要免疫活性成分的单克隆抗体，对其进行检测和定量测定。通过免疫印迹法和酶联免疫吸附法（ELISA），对已建立的小鼠骨髓瘤细胞和经阿玉血清免疫的小鼠脾脏淋巴细胞进行杂交瘤的筛选和克隆，制备单克隆抗体。用鳗弧菌免疫的鳗血清和鳗弧菌脂多糖致敏的聚苯乙烯微球制备抗鳗抗体单克隆抗体。对免疫和未免疫的阿玉进行血清、肾、脾、肝匀浆检测。对某孵化场产的鱼卵和鱼苗也进行了检测。共获得12个单克隆抗体（“A”-“L”）。单克隆抗体“A”被认为是针对ayu抗体的单克隆抗体。血清、肾、脾、肝及薯条匀浆中均检测到抗原“a”-“1”，与单克隆抗体“a”-“L”对应，鸡蛋中仅检测到抗原“b”-“e”和“1”。免疫后的阿育鱼中“a”抗原的浓度高于未免疫的阿育鱼，而其他抗原的浓度未因免疫而增加。