猪抗克伦特罗(CL)抗血清的制备及其IgG纯化与鉴定  

The preparation of pig anti-clenbuterol sera and purification and identification of its serum IgG

作　　者：黄永科[1] 刘清[1] 周光宏[1] 

机构地区：[1]南京农业大学动物科技学院,江苏南京210095

出　　处：《畜牧与兽医》2001年第3期6-8,共3页Animal Husbandry & Veterinary Medicine

摘　　要：为了获得高特异性、高纯度的猪抗克伦特罗 (CL)IgG ,将克伦特罗跟牛血清白蛋白 (BSA)偶联后 ,选取 1 0头大×大×约三元杂交猪 (5头为免疫组 ,5头为对照组 ) ,用偶联物BSA CL对其进行免疫 ,来制备猪抗CL抗血清。以卵清蛋白 (OVA)跟CL的偶联物OVA CL为包被抗原 ,采用ELISA法测定抗血清效价和血清阻断率。结果表明 ,有 4头猪体内产生了抗CL抗体 ,且在第 2次加强免疫后达到最大 ,效价为 1∶2 0 0 0 0 ;而当血清稀释率为 1∶40 0 ,CL的PBS液浓度在 1 8× 1 0 - 4 ～ 7 0× 1 0 - 7之间时 ,其阻断率为 80 %～ 1 8%。随后用正辛酸 硫酸铵法对血清抗体进行纯化 ,经紫外吸收法测定和SDS PAGE电泳试验结果表明IgG成份得到了纯化 。To gain purified special pig anti clenbuterol sera IgG,5 pigs were immunized against clenbuterol(CL)conjugated to bovine sera albumin(BSA)by dizotization.Antibody titers were determined using indirect ELISA with OVA(Ovalbumin) CL conjugated as an antigen in coating the microwells.Indirect inhibitory ELISA was used to detect the anti CL specificity of the sera.The specific anti CL sera were detected in four pigs.Finally the caprylid method was used to purify the immunoglobulin(IgG).The purified IgG was identified by SDS PAGE.The result indicated that the IgG was purified and could be used in further immunization experiment.

关 键 词：猪 克伦特罗 抗克伦特罗抗血清 抗克伦特罗抗体 纯化 鉴定 免疫试验 

分 类 号：S858.282.5[农业科学—临床兽医学]