沙丁胺醇单克隆抗体的制备及其免疫学鉴定

Preparation and Immunologic Characteristics of Monoclonal Antibody for Salbutamol

采用混合酸酐法合成SAL-BSA和SAL-OVA,用紫外法和凝胶电泳法鉴定,使用细胞融合技术建立沙丁胺醇杂交瘤细胞株,通过体内诱生腹水法大量制备沙丁胺醇高亲和力单克隆抗体。结果显示,所免疫的小鼠抗体效价均达到10^-4;融合后筛选出的3株杂交瘤细胞单抗亚型均为IgG1,细胞培养上清间接ELISA效价为1∶2.5×10^2-1∶1.02×10^3,用4D3E7细胞株制作的腹水效价为1∶5.12×10^5,对SAL的IC50为1.35μg/L,除与瘦肉精有10.8%交叉外,与其他β2-受体激动剂的交叉反应均小于0.2%。且具有较好的特异性。试验获得了高效价、敏感、特异的SAL mAb,为动物性食品中SAL残留的免疫学检测奠定了基础。

SAL-BSA and SAL-OVA were produced by the mixed anhydride synthesis and determined with uv method and gel electrophoresis method appraisal,then albuterol hybridoma cell line was established by cell fusion technology,and albuterol high affinity monoclonal antibody was prepared through the body nudged ascites method.The results showed that the mice immune antibody titer reached 10^-4;After the integration screening,three strains were IgG1subtypes of hybridoma L mAb, whose cell culture supernatant had indirect ELISA titer ranging from 1∶2.5×10^2 to 1∶1.02×10^3. Specially,4D3E7cell line had ascites valence 1∶5.12×10^5,SAL IC50of 1.35μg/L,and good specificity for less than 0.2% cross reaction with otherβ2-agonists except for lean meat fine（10.8%）. Therefore,the test obtained high titre,sensitive,specific SAL mAb,which laid a foundation for animal food SAL residual immunology testing.