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以福尔马林灭活的嗜麦芽寡养单胞菌免疫BALB/c小鼠,通过杂交瘤技术制备针对鱼类致病性嗜麦芽寡养单胞菌的单克隆抗体(McAb),用软琼脂法进行克隆化培养,通过间接ELISA法对所得的杂交瘤细胞株进行特异性筛选,并鉴定其Ig亚类,测定腹水效价,进行了相对亲和力和抗原位点等生物学特性分析。结果显示,获得了8株可分泌特异性McAb的杂交瘤细胞,分别命名为1C3、1C6、4H8、3D9、2B11、3G3、2H11、1B8。经过鉴定,这8株McAb能够特异性地针对嗜麦芽寡养单胞菌,与其他测试菌之间无交叉反应;其抗体主要分为4个亚类,分别为IgG2a型、IgG2b型、IgG3型和IgM型;腹水效价均在10-6以上;相对亲和力较高;2B11针对的嗜麦芽寡养单胞菌脂多糖表位,而其余7株McAb针对的非脂多糖抗原位点。试验结果证实成功制备了能稳定分泌抗嗜麦芽寡养单胞菌McAb的细胞株,为水产养殖中该菌的快速检测和免疫学研究奠定了基础。
BALB / c mice were immunized with formalin-killed Stenotrophomonas maltophilia and monoclonal antibodies (McAbs) against pathogenic Stenotrophomonas maltophilia were prepared by hybridoma technique. The clones were harvested by agar method. The hybridoma cell lines were screened by indirect ELISA and their Ig subclasses were identified. The ascites titer was determined. The biological characteristics such as relative affinity and antigenic site were analyzed. The results showed that eight hybridoma cells secreting specific McAb were obtained and named 1C3, 1C6, 4H8, 3D9, 2B11, 3G3, 2H11 and 1B8 respectively. After identification, these 8 strains of McAbs could specifically target Stenotrophomonas maltophilia and had no cross-reaction with other tested bacteria. The McAbs were divided into 4 subclasses: IgG2a, IgG2b and IgG3 And IgM type; ascites titer above 10-6; higher relative affinity; 2B11 against Stenotrophomonas malt lipopolysaccharide epitopes, while the remaining seven McAb against non-lipopolysaccharide antigen sites. The test results confirmed that the cell strain that can stably secrete McAb against Stenotrophomonas maltophilia was successfully prepared, which lays the foundation for the rapid detection and immunological study of the bacteria in aquaculture.