【摘要】：目的制備并鑒定2型豬鏈球菌(Streptococcus suis serotype 2,S.suis 2)四川分離菌株05ZYH33的溶血素(Suilysin,SLY)多克隆和單克隆抗體。方法利用重組SLY蛋白制備SLY兔多抗,優(yōu)化間接ELISA體系檢測多抗效價(jià),Western blot分析SLY多抗特異性,檢測SLY兔多抗對(duì)人外周血?dú)⒕饔玫挠绊�。利用雜交瘤細(xì)胞融合技術(shù)制備SLY單抗,篩選雜交瘤細(xì)胞株并制備小鼠腹水,間接ELISA和Western blot鑒定SLY單抗的效價(jià)和特異性。結(jié)果間接ELISA顯示SLY多抗效價(jià)高達(dá)1∶204 800,Western blot顯示SLY多抗有較高的特異性和免疫反應(yīng)性,人外周血?dú)⒕囼?yàn)顯示,SLY多抗能加強(qiáng)人外周血對(duì)05ZYH33的殺傷作用。間接ELISA檢測雜交瘤細(xì)胞2B6和5F7培養(yǎng)上清效價(jià)分別是1∶800和1∶3 200,單抗效價(jià)都為1∶204 800。間接ELISA和Western blot鑒定顯示,單抗2B6和5F7具有很強(qiáng)的特異性和免疫反應(yīng)性。結(jié)論成功制備了效價(jià)高、特異性強(qiáng)的S.suis 2 05ZYH33 SLY多克隆和單克隆抗體。
[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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