抗鱼类致病性嗜麦芽寡养单胞菌单克隆抗体的制备及其生物学特性鉴定  

Preparation and characterization of monoclonal ant ibodies against pathogenic Stenotrophomonas maltophilia in fish

作　　者：黄艺丹[1] 汪开毓[1] 郑建[2,3] 黄锦炉[1] 

机构地区：[1]四川农业大学鱼病研究中心,四川雅安625014 [2]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016 [3]Artron Bio Research公司,加拿大温哥华V5J5H6

出　　处：《中国兽医科学》2010年第9期907-912,共6页Chinese Veterinary Science

基　　金：农业部动物检疫检测与防治项目(2130108);中央级农业公益性行业科研专项经费项目(200803013);教育部“长江学者和创新团队发展计划”创新团队项目(IRT0848);加拿大合作项目(Artron BioResearch公司2007)

摘　　要：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 Stenotrophomonas maltophilia ina ctivated by formalin.The hybridoma cell lines consistently secreting monoclonal antibodies（McAbs）against pathogenic S.maltophilia in fish were obtained through cell fusion.The specificity of the McAbs was analysed by an indirect ELISA.Then the subtypes were identified,the titer and relative affinity were measured,and its specifici ty was analyzed.Examination were made to identify the antigen epitope of McAbs by combining expe rimental site and indirect ELISA with lipopolysaccharides（LPS）of S.mal tophilia.In results,from over hundreds of positive hybridomas which secreted anti-S.maltophilia McAbs,eight strains of hybridomas were screened out,and designated as 1C3,1C6,4H8,3D9,2B11,3G3,2H11 and 1B8,respectively.The subtypes of the McAbs were IgG2a,IgG2b,IgG3 and IgM,respectively.The titers of these McAbs produced by ascites fluid were higher than 10-6.The McAbs had high relativ e affinity.The eight strains of McAbs targeted different antigen epitope：only 2B 11 targeted the LPS of S.maltophilia,while the others targeted t he non-LPS sites of S.maltophilia.In conclusion,eight specific McAbs against S.maltophilia were ob tained successfully,which might be very useful for further studies of the patho genesis and early diagnosis of S.maltophilia in aquaculture.

关 键 词：嗜麦芽寡养单胞菌 单克隆抗体 生物学特性 

分 类 号：S947.12[农业科学—水产养殖] S852.614[农业科学—水产科学]