2型猪链球菌溶血素多克隆和单克隆抗体制备及鉴定

发布时间：2019-05-19 11:00

【摘要】：目的制备并鉴定2型猪链球菌(Streptococcus suis serotype 2,S.suis 2)四川分离菌株05ZYH33的溶血素(Suilysin,SLY)多克隆和单克隆抗体。方法利用重组SLY蛋白制备SLY兔多抗,优化间接ELISA体系检测多抗效价,Western blot分析SLY多抗特异性,检测SLY兔多抗对人外周血杀菌作用的影响。利用杂交瘤细胞融合技术制备SLY单抗,筛选杂交瘤细胞株并制备小鼠腹水,间接ELISA和Western blot鉴定SLY单抗的效价和特异性。结果间接ELISA显示SLY多抗效价高达1∶204 800,Western blot显示SLY多抗有较高的特异性和免疫反应性,人外周血杀菌试验显示,SLY多抗能加强人外周血对05ZYH33的杀伤作用。间接ELISA检测杂交瘤细胞2B6和5F7培养上清效价分别是1∶800和1∶3 200,单抗效价都为1∶204 800。间接ELISA和Western blot鉴定显示,单抗2B6和5F7具有很强的特异性和免疫反应性。结论成功制备了效价高、特异性强的S.suis 2 05ZYH33 SLY多克隆和单克隆抗体。
[Abstract]:Objective to prepare and identify hemolysin (Suilysin,SLY) polyclone and monoclonal antibody against Streptococcus suis type 2, S.suis 2) strain 05ZYH33 isolated from Sichuan. Methods SLY rabbit polyantibodies were prepared by recombinant SLY protein. The specificity of SLY polyantibodies was analyzed by, Western blot with optimized indirect ELISA system, and the effect of SLY rabbit polyantibodies on the bactericidal effect of SLY rabbits on human peripheral blood was detected. Hybridoma cell fusion technique was used to prepare SLY monoclonal antibody, hybridoma cell line was screened and mouse ascitic fluid was prepared. Indirect ELISA and Western blot were used to identify the titer and specificity of SLY monoclonal antibody. Results indirect ELISA showed that the titer of SLY polyclonal antibody was as high as 1 鈮，

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