摘要
将氯霉素(CAP)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氯霉素半琥珀酸酯(CAP-HS);采用混合酸酐(MA)法将CAP-HS与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联合成人工免疫原BSA-CAP-HS和包被原OVA-CAP-HS,用红外(IR)、紫外(UV)、凝胶电泳(SDS-PAGE)进行鉴定;用BSA-CAP-HS免疫BALB/C小鼠,间接ELISA测定多抗(pAb)效价,阻断ELISA鉴定其敏感性,交叉反应试验鉴定其特异性。结果表明,CAP-HS与BSA合成成功,其分子结合比为15.6∶1,获得了高价、敏感、特异的CAP pAb,为CAP残留免疫学检测方法的建立奠定了基础。
The active group carboxyl was introduced to Chloramphenicol（CAP） and formed CAP half succinaate（CAP-HS） which has hapten structure by chemical modification. The method of mixed anhydride（MA） was used to conjugate CAP-HS to BSA and artificial immunogen BSA-CAP-HS was obtained, the coating antigen OVA-CAP-HS was obtained in the same way. IR, UV and SDS-PAGE were used to identify BSA-CAP-HS. BALB/C mice were immunized with BSA-CAP-HS, the titer of polyclonal antibody（pAb） was detected by indirect ELISA, the sensitivity of pAb was identified by blocking ELISA and the specificity of pAb was identified by cross-reaction test. The results showed that BSA-CAP-HS was synthesized successfully and its conjugation ratio of CAP-HS to BSA was about 15.6： 1, the high-titer, sensitive and specific CAP pAb were preduced in sera of immunized BALB/C mice, which made it possible to establish immybiassay of CAP residues in feed and animal food.
出处
《安徽农业科学》
CAS
北大核心
2006年第9期1751-1753,1757,共4页
Journal of Anhui Agricultural Sciences
基金
国家"863"项目(2001AA249030)资助
关键词
氯霉素
半抗原
人工免疫原
合成
鉴定
Chloramphenicol
Hapten
Artificial immunogen
Synthesis
Identification
