抗鸡IgM单克隆抗体的研究──Ⅱ．检测抗鸡IgMμ链单克隆抗体的间接ELISA法  

Research on Monoclonal Antibodies Specific to Chicken IgM ──Ⅱ. Detection of Monoclonal Antibody against μ-chain of Chicken IgM by indirect Enzyme-linked Immunosorbent Assay

作　　者：钱建飞[1] 毛洪先[1] 邵国青[1] 金洪效[1] 

机构地区：[1]江苏省农业科学院畜牧兽医研究所

出　　处：《江苏农业学报》1994年第3期29-33,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金;江苏省科委"八五"课题

摘　　要：采用饱和硫酸铵盐析、ＳｅｐｈａｄｅｘＧ２００柱层析和羊抗鸡ＩｇＭμ链抗血清亲和层析等方法从鸡血清中纯化ＩｇＭ，同时采用硫酸钠盐析及ＤＥＡＥ离子交换层析从鸡蛋黄中纯化ＩｇＧ。以纯化的ＩｇＭ和ＩｇＧ包被酶标反应板，进行双排筛选，建立了检测抗鸡ＩｇＭμ链单克隆抗体的间接ＥＬＩＳＡ法。ＩｇＭ和ＩｇＧ最适包被浓度为１μｇ／ｍｌ，ＨＲＰ－羊抗鼠ＩｇＧ最适工作浓度为１：２０００。从４７８孔杂交瘤细胞中共检出３０孔阳性，阳性检出率约为６．２８％。试验结果表明此方法具有特异性强、重复性好、敏感性高等优点。his project is suppported by the National Natural Science Fuondation of China and by the Commission of Science and Technology of Jiangsu Prouince. Chicken IgM was purified from serum by a procedure of precipitation with saturated ammonium sulfate, chromatography with Sephadex G200 and affinity chromatography with goat anti-chicken IgM μ-chain. Chicken IgG was purified from egg yolk by precipitation with natrium sulfate and ion-exchange chromatography with DEAE. An indirect enzyme-linked immunosorbent assay (ELISA ) for detection of monoclonal antibody (McAb ) against μchain of chicken IgM was established by the plastic plate for ELISA coated with purified chicken IgM and IgG. The optimum concentrations were 1 μg/ml IgM and IgG for coating and 1: 2000 dilution of HRP-goat anti-mouse IgG for working. Thirty positive hybridoma wells were screened from 478 hybridoma wells and the positive rate was about 6. 28%. The experimental results snow that this indirect ELISA for detection of McAb specific to μ-chain of chicken IgM is specific, sensitive and repeatable.

关 键 词：鸡IgM 单克隆抗体 间接ELISA 

分 类 号：S858.312.4[农业科学—临床兽医学]