恩诺沙星完全抗原的制备与鉴定及其多克隆抗体的制备

摘要

通过活化酯法将小分子的恩诺沙星（EFLX）偶联到牛血清白蛋白（BSA）和鸡卵清白蛋白（OVA）上，形成偶联物EFLX-BSA和EFLX-OVA，以EFLX-BSA为免疫原试验小鼠，以EFLX-OVA为检测原通过ELISA检测小鼠血清效价，以及Ci-ELISA检测抗体的特异性和最低检测限。结果表明免疫原和检测原均成功合成，血清效价检测结果第一组为25600，第二组和第三组均大于51200，第二组最高；间接竞争ELISA法结果表明，所获抗体对EFLX特异性强，最低检出限为1．17 ng/mL，IC50值为54．60 ng/mL，检测范围为7．39～403．43 ng/mL。本试验所获得抗体效价高、特异性强，可以为ELISA试剂盒和胶体金试纸条研发提供良好的抗体材料。%Ovalbumin ( OVA) and bovine serum albumin ( BSA) were used as protein carriers to synthesize small molecule drugs EFLX complete antigens by the active ester method .The conjugates formed in this way were EFLX-BSA and EFLX-OVA.The serum titer was detected by means of EFLX-BSA immunogen and EFLX-OVA coating antigen by ELISA ,the specificity and detection limit ( LD) were detected by Ci-ELISA.The results showed that the immunogen and coating antigen were successfully synthesized .The serum titer detection result of the first group was 25 600,those of the second and third groups were more than 51 200,that of the second was the highest of all .The result of Ci-ELISA indicated that this method was sensitive and had a linear detec-tion range from 7.39 ng/mL to 403.43 ng/mL ( R2=0.993 9) ,with IC50 and LOD values of 54.60 ng/mL and 1.17 ng/mL.This experiment obtained high antibody titer and the specificity was strong ,which provides good antibody materials for the development of colloidal gold test and ELISA kit .