抗传染性犬肝炎病毒单克隆抗体的制备及鉴定

Preparation and Characterization of Monoclonal Antibody against ICHV

目的制备传染性犬肝炎病毒单克隆抗体。方法将用纯化的犬腺病毒1型（CAV-1）免疫的BALB／c小鼠脾细胞与SP2／0细胞在聚乙二醇作用下融合，通过酶联免疫吸附试验（ELISA）和免疫酶试验（1EA）筛选，以有限稀释法克隆3次，制备单克隆抗体，并对制备完成的单克隆抗体进行生物学鉴定。结果成功得到2株能稳定分泌抗CAV-1的单克隆抗体杂交瘤细胞，命名为1B、2H3，经鉴定其亚型分别为IgG2a和IgG2a，2株均为kappa链。腹水的ELISA效价可达10^7-10^8，IEA效价可达1：2560—1：5120。该单克隆抗体与CPV、CDV、FPV、CCV病毒无交叉反应。结论成功制备了抗CAV-1单克隆抗体，为进一步建立相关诊断方法奠定了基础。

Objective To prepare and identify the monoclonal antibody against Infections Canine Hepatitis virus. Methods BALB/c mice were immunized conventionally with the purified CAV-1. On the third day after the final immunization, spleen cells of mice fused with myeloma cells SP2/0 were filtered by culturing selectively, detecting special antibody, ELISA test, IEA and coloning. Two hybridoma cell lines, obtained by culturing in vitro and anabiosis after freezing in a long period which were named lET ,2H3 ,could stably secrete McAbs. Results The subclasses of the two McAbs were IgG2a and IgG2b respectively, Kappa light chain. The ELISA and IEA titers of aseites were 1 ： 107 - 1 ： l0s , 1 ： 2560 - 1 ： 5120 respectively. These two McAbs were specific to CAV-1, but not reacted with CPV, CDV, FPV, CCV. Conclusion McAb against CAV-1 has been successfully prepared. It is also a basic for establishing a related diagnoses method.