大黄鱼鼠多抗浅析

摘 要 IgM 是大黄鱼最主要的免疫球蛋白，在鱼体免疫系统中发挥重要作用。本论文以实验室前期利用 Protein A 亲和纯化的大黄鱼血清 IgM 为抗原，按照标准免疫流程免疫 Balb/C 小鼠，获取抗血清，即多克隆抗体。利用酶联免疫吸附方法（ELISA）测定鼠源抗血清的效价达到 1:20000，效果较好；利用免疫印迹（Western blotting）对大黄鱼血清 IgM 鼠源多抗的结合特性进行了分析，结果显示多抗能特异性结合 IgM 重链和轻链，表明实验制备的多抗特异性良好，可应用于后续实验研究。大黄鱼血清 IgM 鼠多抗的制备有助于加深对鱼体免疫系统的理解，也将为大黄鱼免疫机理及相关免疫检测研究奠定基础。关键词：大黄鱼；IgM；鼠多抗；ELISA；Western blottingAbstractIgM is the main immunoglobulin in large yellow croaker, which plays a vital role in the immune system of fish. In this thesis, the serum IgM of Larimichthys crocea purified by protein A affinity, according to the standard immune process, was used as antigen to immunize Balb/c mice, and the antiserum, polyclonal antibody, was acquired. The valence of mouse antiserum determined by enzyme-linked immunosorbent assay (ELISA) achieved 1:20000. Western blotting was used to analyze the binding characteristics of murine polyclonal antibodies against serum IgM of L. crocea. The results showed that the polyclonal antibodies could specifically combine with heavy and light chains of IgM L. crocea, which indicated that it prepared in this experiment had good specificity and could be applied to subsequent experimental studies. The preparation of rat polyclonal antibody against the serum IgM of L. crocea helps to understanding of the immune system of the fish, and will also lay a foundation for the study of the immune mechanism and related immune detection of L. crocea.Key words： Larimichthys crocea; IgM; mice polyclonal antibody; ELISA; Western blotting目 录1 引 言..........................