沙丁胺醇人工抗原的合成与鉴定

通过沙丁胺醇(SAL)与琥珀酸酐的醇解反应制备SAL的琥珀酸衍生物,用EDC法将SAL琥珀酸衍生物偶联于载体蛋白BSA和OVA,合成免疫原BSA-SAL和包被原OVA-SAL,用紫外扫描(UV)、SDS-PAGE进行鉴定;用BSA-SAL免疫BALB/C小鼠,间接ELISA测定多抗上清(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,BSA-SAL的UV与BSA、SAL的相比发生了改变;通过UV分析,SAL-HS与BSA的分子结合比约为7.2∶1;通过系列ELISA鉴定,获得了高价、敏感、特异的pAb,为SAL残留免疫学检测方法的建立奠定了基础。

Synthesis and identification of salbutamol artificial antigen

Salbutamol(SAL) succinate was prepared by means of the alcoholysis reaction between SAL free base and succinic anhydride.SAL succinate was conjugated to carrier protein BSA by the method of EDC,and thus the antigen BSA-SAL for immunization was obtained.The coating antigen OVA-SAL was obtained in the same way.UV and SDS-PAGE were used to identify SAL artificial antigen.BALB/C mice were immunized with BSA-SAL,the titre 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 the method of cross-reaction test.The results showed that compared with BSA and SAL,UV spectrogram of BSA-SAL has changed;the conjugation ratio of SAL-HS to BSA was about 7.2∶1;the high-titer,sensitive and specific anti-SAL pAb has been produced in sera of immunized BALB/C mice,which made it possible to establish immunoassay of SAL residues in feed and animal food.