摘要
利用小鼠骨髓瘤Sp2/0细胞系与纯化鸡IgG免疫的BALB/C小鼠脾细胞融合,成功地建立了5株能持续分泌抗鸡IgG单克隆抗体(McAbs)的杂交瘤细胞株。选择产生抗体效价高、生长快的C_(44)株,扩大培养后注射于小鼠制备腹水,经硫酸铵盐析粗提后,用HRP标记,制备出McAbs酶结合物。将制备的McAbs酶结合物和多克隆抗体(PcAbs)酶结合物,采用间接ELISA,对35份鸡血清中鸡新城疫病毒(NDV)抗体以及19份鸡血清中鸡嗜血杆菌(HG)抗体进行检测,McAbs和PcAbs两者的相关系数分别为0.928和0.921,证明制备的抗鸡IgG McAbs具有特异性强、效价高的特点.
Five hybridomas secreting anti-chicken IgG McAbs wereestablished by the fusion of mouse myeloma cells Sp 2/0 and spleencells from BALB/C mouse immunized with purified chicken IgG.The culture supernates were assayed for the presence of antibody-producing cells by ELISA, and their specificities were identified bydouble immunodiffusion test, indirect and sandwich ELISA. Miceinoculated intraperitoneally with the cloned hybridomas could pro-duce McAbs in the ascitic fluid. The HRP-labelled McAbs and HRP-labelled PcAbs were both separately used for detecting serumantibodies of Newcastle disease virus (NDV) and Haemophilusgallinarum (HG) by indirect ELISA. The correlation coefficientwere 0. 928 and 0.921 in the antibody detection of NDV and HGrespectively. It is considered that the McAbs could become a newreagent for chicken disease diagnosis and supervision.
