摘要:
亲和层析纯化鳜鱼血清免疫球蛋白,制备针对鳜鱼血清免疫球蛋白的兔多克隆抗体,采集常见经济鱼类血清25种,利用优球蛋白法纯化血清,通过间接ELISA的方法筛选与兔抗鳜鱼Ig反应的鱼类血清,结合变性还原条件与非变性非还原条件下的蛋白印迹试验显示:制备的兔抗鳜鱼Ig识别鲈形目中鳜鱼、加州鲈鱼、卵形鲳鲹、尼罗罗非鱼、花鲈、红罗非鱼、青石斑鱼、大黄鱼、黑鲷、斜带髭鲷、美国红鱼,鲽形目中的牙鲆、大菱鲆,鮋形目中的褐菖鮋血清免疫球蛋白的重链与轻链。而与鲤形目中的5种鱼,鲇形目中的4种鱼,鳗鲡目中的海鳗、欧洲鳗鲡反应条带较为复杂。嗜水气单胞菌外膜蛋白免疫刺激复合物(ISCOMs)免疫尼罗罗非鱼,间接ELISA 法检测免疫组血清中的抗体,利用兔抗鳜鱼Ig多克隆抗体作为二抗,结果显示,与对照组相比,免疫组有较高的抗体水平。
关键词:
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鱼类 /
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血清 /
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Ig /
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多克隆抗体 /
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蛋白印迹
Abstract:
Siniperca chuatsi serum Ig was purified by affinity chromatography for prepared polyclonal antibody against Siniperca chuatsi serum Ig.Twenty-five fish's sera were collected and purified the serum Ig by euglobulin precipitation.Serum Ig could react with polyclonal antibody that against Siniperca chuatsi serum Ig were screened.The polyclonal antibody could recognize Siniperca chuatsi,Micropterus salmoides,Trachinotus ovatus,Oreochromis niloticus,Lateolabarax japonicus,Oreochromis sp., Epinephelus awoara,Pseudosciaena crocea, Sebastiscus marmoratus, Sciaenops ocellatus, Sparus macrocephalus, Scophthatmus maximus, Paralichthys olivaceus, Sebastiscus marmoratus, serum Ig heavy chain and light chain.The polyclonal antibody reacted with many protein bands from purified serum of five species Cypriniformes, four species fish of Siluriformes, purified serum of Anguilla anguilla and Muraenesox cinereus.Oreochromis niloticus were immunized intraperitoneally with aeromonas hydrophilla ZN1 outer membrane proteins immune stimulating complexes(ISCOM).The antibody in the serum was detected by ELISA, the polyclonal antibody against Siniperca chuatsi from rabbit was used as second antibody.The result showed that the antibody from immunized group was higher than that from control group.
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