分泌抗山羊γ干扰素单克隆抗体杂交瘤细胞株的制备与鉴定

Preparation and Identification of Hybridoma Cell Lines Secreting Antibodies against Goat IFN-gamma

为筛选分泌山羊γ干扰素(IFN-γ)单克隆抗体的杂交瘤细胞,以原核表达的山羊rIFN-γ蛋白免疫Balb/c小鼠,取其脾脏细胞与SP2/0瘤细胞进行细胞融合,以间接ELISA方法筛选分泌山羊IFN-γ单克隆抗体杂交瘤细胞,采用山羊外周血淋巴细胞产生的IFN-γ包被酶标板对IFN-γ的特异性进行鉴定,用试纸条对IFN-γ单克隆抗体类别和亚型进行鉴定。结果获得了1株能特异性识别山羊IFN-γ的杂交瘤细胞3C,3C分泌的单克隆抗体能够特异性识别天然结构的山羊IFN-γ,单克隆抗体类别为IgG,轻链为κ型。

In order to prepare hybridoma cells secreting monoclonal antibodies against goat IFN-γ,Balb/c mice were immunized with purified prokaryotic reeombiant goat IFN-γ（rIFN-7） and the splenocytes were isolated and fused with SP2/0 myeloma to generate hybridoma cells.Then the supernatant was collected to identify the monoclonal antibodies recognizing goat IFN-γ specifically. In the present study, an I-EI.ISA method was established to detect antibodies against goat IFN-γ when the antigen was coated with goat lymphocytes.Then the classes and types of monoclonal antibodies against goat IFN-γ were characterized by quick test strip.The results suggested that a strain of hybridoma cell named 3C which showed high specificity was prepared successfully,and the monoclonal antibody was identified as IgG class and kappa type.