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当前位置: 首页 > 文章 > “新吉富”罗非鱼免疫球蛋白单克隆抗体的制备及其在免疫效果评价上的应用 福建农林大学学报(自然科学版) 2017,46 (2) 187-191
Position: Home > Articles > Preparation of Mc Abs against immunoglobulinin from Oreochromis niloticus and its application in immune effect analysis Journal of Fujian Agriculture and Forestry University(Natural Science Edition) 2017,46 (2) 187-191

“新吉富”罗非鱼免疫球蛋白单克隆抗体的制备及其在免疫效果评价上的应用

作  者：
吴斌
单  位：
福建省水产技术推广总站;福建省淡水水产研究所
关键词：
新吉富罗非鱼;免疫球蛋白IgM;单克隆抗体;ELISA检测方法
摘  要：
制备了"新吉富"罗非鱼血清免疫球蛋白IgM单克隆抗体(McAb)并进行了特性分析,以血清IgM McAb为基础,建立了IgM捕获ELISA检测方法.采用亲和层析法纯化健康"新吉富"罗非鱼IgM,纯化蛋白经SDS-PAGE检测,重链、轻链的相对分子质量分别为75~78、23~25 ku;以纯化的IgM为抗原制备IgM McAb,制备McAb杂交瘤细胞株系3株,分别命名为2H5、2D4、2B11,抗体亚型均为IgM,小鼠腹水效价分别为1.0×10~(-5)、1.0×10~(-4)、1.0×10~(-5),对IgM敏感度测定表明,2H5、2D4、2B11检测灵敏度分别为31.25、125.00、62.50 ng.以抗罗非鱼IgM McAb包被酶标板,建立IgM捕获ELISA检测方法.以无乳链球菌灭活疫苗免疫罗非鱼,受免血清按建立的IgM捕获ELISA方法检测特异性抗体,结果表明,建立的IgM捕获ELISA检测方法可应用于免疫应答抗体水平分析.
译  名：
Preparation of Mc Abs against immunoglobulinin from Oreochromis niloticus and its application in immune effect analysis
作  者：
WU Bin;Fujian Provincial Fishery Technical Extention Center/Freshwater Fisheries Research Institute of Fujian;
关键词：
Oreochromis niloticus;;immunoglobulin;;monoclonal antibody;;ELISA
摘  要：
To prepare effective monoclonal antibody( McAb) against serum immunoglobulin( Ig) of tilapia,antibody capture enzyme-linked immunosorbent assay( ELISA) was conducted,and Ig of healthy Oreochromis niloticus was purified by protein A affinity chromatography to characterize its immunological feature. Then purified IgM was used as antigen to immunize Balb/c mice. SDSPAGE analysis showed that molecular weight of tilapia Ig heavy chain and light chain was 75-78 and 23-25 ku,respectively. Three hybridomas strains secreting monoclonal antibodies( McAb) were obtained and named 2H5,2D4,and 2B11,with the common isotype being IgM. And titers in the ascites of Balb/c mice were 1.0× 10~(-5),1.0×10~(-4),1.0×10~(-5),respectively. Sensitivities of McAb against tilapia IgM was high,with detection limits being 31. 25,125. 00,and 62. 50 ng,respectively. Furthermore,MAC-ELISA showed that specific antibody was detected within tilapia immuned by Streptococcus agalaciate inactivated vaccine.

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