鸡IgG单克隆抗体的制备与鉴定

目的]制备鸡免疫球蛋白单克隆抗体,提高诊断鸡特异性抗体的水平。方法]应用盐析法粗提和葡聚糖G200凝胶层析分离纯化鸡血清IgG,免疫BALB/c小鼠,并进行细胞融合和ELISA筛选。结果]间接ELISA法检测C44、C45、C67和C68共4株鸡IgG单克隆抗体的杂交瘤细胞的腹水效价分别为1∶640000、1∶320000、1∶640000和1∶80000。Westernblot分析显示,C44和C45株单克隆抗体识别鸡IgG轻链,而C67和C68株单克隆抗体识别鸡IgG重链,它们与鸭、猪等血清Ig均没有反应。小鼠Ig亚类分析表明,4株细胞分泌的抗体均为IgG1型。结论]成功获得了4株稳定分泌鸡IgG单克隆抗体的杂交瘤细胞。

Production and Identification of Monoclonal Antibody against Chicken IgG

Objective]To preparate monoclonal antibody against chicken Ig and improve the diagnostic level of specific antibodies in chickens.Method]Firstly,chicken IgG was isolated by saturated ammonium sulfate and purified by DEAE G200; secondly,BALB/c mice were immunized with chicken IgG and a cell fusion of the spleen and SP2/O strain was carried out,thirdly,positive samples were detected and selected with ELISA.Result]Four strains secreting monoclonal antibody were obtained and named as C44,C45,C67 and C68.Their titers of ascites in ELISA were 1∶640 000,1∶320 000,1∶640 000 and 1∶80 000,respectively.C44 and C45 recognized chicken IgG light chain and C67 and C68 recognized heavy chain.They all could not recognize Ig from duck,rabbit and swine.Additionally,the Ig type identification results showed that they all belonged to IgG1.Conclusion]Four strains of obtained hybridoma can stably product the monoclonal antibody against chicken Ig.