氯霉素人工免疫原的合成与鉴定

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。

Synthesis and Identification of Chloramphenicol Artificial Immunogen

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 OVACAP-HS was obtained in the same way.IR,UV and SDS-PAGE were used to identify BSACAP-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 produced in sera of immunized BALB/C mice,which made it possible to establish immunoassay of CAP residues in feed and animal food.