高亲和力抗IBDV单克隆抗体的产生与免疫学鉴定

Preparation and identification of high affinity monoclonal antibodies to IBDV

目的：生产高亲和力的抗IBDV不同抗原决定簇的单克隆抗体，为IBD的快速检测准备试剂。方法：用IBDV抗原诱导Balb/c小鼠产生最强最适当的反应，将免疫脾细胞与NSO瘤细胞融合，通过对大量的杂交细胞培养上清进行检测和鉴定，筛选出分泌高亲和力单克隆抗体的杂交瘤细胞。结果：获得分泌高亲和力单克隆抗体的杂交瘤21株，其中4株YNZ-1、YNZ－12、YNZ-8和YNZ-26的间接ELISA效价分别为：1：2560、1：1280、1：2560、1：640；腹水间接ELLSA效价分别为：1×10－6、5×10－5、1×10－6和1×10-5。结论：单抗YNZ－1和YNZ-18、YNZ－12和YNZ－26亲和力高，识别IBDV不同的抗原决定簇，可作为制备快速诊断IBD的特异单克隆抗体。

Objective: Preparation of high affinity monoclonal antibodies to different epitopes on IBD virus. Methods: Spleen cells from Balb/c mice immunized with IBDV antigen were fused with NSO myeloma cell line and the supernatant was screened for specific monoclonal antibodies with high affinities. Results: Twenty-one hybridoma lines secreting monoclonal antiodies specific to IBDV were selected and cloned for 30 fusions. Four of the lines were identifiec to secret antibodies to different epitopes with high affinity. Indirect ELISA titers of the four clones,YNZ-1, YNZ-12,YNZ-18 and YNZ-26, were as the following: 1:2 560, 1: 1 280, 1:2 500, 1:640(supernatant), 1×10-6, 5×10-3, 1×10-6 and 1×10-5 (AF) .Conclusion: Monoclonal antibodies YNZ-1 and YNZ-18 recognize one epitope on the IBDV whereas YNZ12 and YNZ-26 identify another.