大黄鱼免疫球蛋白IgM鼠源多克隆抗体的制备及其结合特性  

Development and Characterization of Mouse Polyclonal Antibodies to Serum IgM in Large Yellow Croaker Larimichthys crocea

作　　者：何亮银 吴建绍[2] 陈卯森 黄伟卿 韩坤煌 李进寿 史晓丽 HE Liangyin;WU Jianshao;CHEN Maosen;HUANG Weiqing;HAN Kunhuang;LI Jinshou;SHI Xiaoli(College of Life Science,Ningde Normal University,Ningde 352100,China;Key Laboratory of Cultivation and High-value Utilization of Marine Organisms in Fujian Province,Fisheries Research Institute of Fujian Province,Xiamen 361013,China;Engineering Research Center of Mingdong Aquatic Product Deep-Processing,Ningde 352100,China)

机构地区：[1]宁德师范学院生命科学学院,福建宁德352100 [2]福建省水产研究所,福建省海洋生物增养殖与高值化利用重点实验室,福建厦门361013 [3]闽东水产品精深加工福建省高等学校工程研究中心,福建宁德352100

出　　处：《水产学杂志》2022年第4期32-37,共6页Chinese Journal of Fisheries

基　　金：福建省海洋生物增养殖与高值化利用重点实验室开放课题基金资助项目(2019fjscq05);宁德师范学院引进人才科研经费资助项目(2017Y03);福建省科技厅自然科学基金项目(2020J01426).

摘　　要：采用硫酸铵盐析结合Protein A亲和层析,分离纯化了体质量300~400 g大黄鱼(Larimichthys crocea)血清免疫球蛋白IgM,制备了鼠源IgM抗血清,并分析了其结合特性。SDS-PAGE结果显示,终浓度为50%的硫酸铵粗提结合Protein A亲和层析获得的IgM纯度高,由2条分子量约为75 kDa和28 kDa的重链和轻链条带组成。酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定鼠源IgM多克隆抗体的效价为20000,免疫印迹(Western blotting)显示了抗血清与IgM的良好结合活性。大黄鱼血清IgM多抗的制备,将为鱼体免疫机制研究及病害防治提供有力工具。In order to investigate the characteristics of immunoglobulins M(IgM)in large yellow croaker(Larimichthys crocea),serum IgM was isolated from large yellow croaker by combination of ammonium sulfate salting out and protein A affinity chromatography,then the purity and molecular weight of the serum IgM were determined by SDS-PAGE.It was found that the purified IgM consisted of two main bands heavy chain and light chain with the molecular weight of 75 kDa and 28 kDa,respectively.Furthermore,healthy mice were immunized with the purified IgM.The ELISA indicated that the titer of antisera was determined to be 20000,and Western blotting revealed the specifical reorganization to IgM.The findings provide a reference for research on IgM characteristics,and the antisera obtained can be used in treatment of large yellow croaker disease caused by pathogen.

关 键 词：大黄鱼 血清IGM Protein A 多克隆抗体 结合特性 

分 类 号：S917.4[农业科学—水产科学]