莱克多巴胺人工免疫原合成及抗体特性  

Studies on Synthesis of Artificial Immunogen and Characteristics of Polyclonal Antibody of Ractopamine

作　　者：张海棠[1] 王自良[1] 王艳荣[1] 邓瑞广[2] 

机构地区：[1]河南科技学院动物科学学院,河南新乡453003 [2]河南省动物免疫学重点实验室,河南郑州450002

出　　处：《西北农业学报》2008年第6期1-5,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：“十一五”国家科技支撑计划重大项目“农药、兽药抗体库建立”资助(2006BAK02A21/1)

摘　　要：将莱克多巴胺(RAC)进行化学修饰引入羧基活性基团,合成具有半抗原结构特征的RAC-戊二酸酐半醛化合物(RAC-SA);采用混合酸酐法(MA)将RAC-SA与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联合成人工免疫原BSA-RAC和包被抗原OVA-RAC,用红外(IR)、紫外(UV)和凝胶电泳(SDS-PAGE)进行鉴定,推算分子结合比;用BSA-RAC免疫新西兰白兔,间接ELISA测定多克隆抗体(pAb)效价,阻断ELISA鉴定其敏感性,琼脂双扩散试验、WestGold膜杂交试验和交叉反应试验鉴定其特异性。结果表明,BSA-RAC偶联成功,分子结合比为24.5∶1;获得了高价、敏感、特异的RAC pAb,为RAC残留免疫学检测方法的建立奠定了基础。The active group carboxyl was introduced to Ractopamine （RAC） and formed RAC semialdehyde （RAC-SA） which has hapten structure by chemical modification. The method of mixed anhydride （MA） was used to conjugate RAC-SA to bovine serum albumin （BSA） and artificial immunogen BSA-RAC was obtained, the coating antigen OVA-RAC was obtained in the same way. Infrared （IR）, ultraviolet （UV） and sodium dodecyl sulfate polyacrylamide gel electropheresis （SDS-PAGE） were used to identify BSA-RAC. New Zealand white rabbits were immunized with BSA-RAC, the titre of polyclonal antibody （pAb） was detected by indirect EI.ISA, the sensitivity of pAb was identified by blocking ELISA and the specificity of pAb was identified by test of agar bidirectional pervasion, WestGold immunochromatography and cross-reaction. The results showed that BSA-RAC was syn thesized successfully and its conjugation ratio of RAC to BSA was about 24.5 ： 1, the high-titer, sensitive and specific RAC pAb were produced in sera of immunized New Zealand white rabbits, which made it possible to establish immunoassay of RAC residues in animal food.

关 键 词：莱克多巴胺 半抗原 人工免疫原 抗体 特性 

分 类 号：S859.84[农业科学—临床兽医学]