二氟沙星抗体的制备

Production of Difloxacin Antibody

[目的]制备二氟沙星(DIF)抗体。[方法]采用N-羟基琥珀酰亚胺活性酯(NHS)法,将二氟沙星分别与牛血清白蛋白(BSA)、卵清白蛋白(OVA)偶联,制备免疫抗原DIF-BSA和检测抗原DIF-OVA,并用DIF-BSA免疫小鼠以获得高效价的抗体。[结果]免疫抗原DIF-BSA的紫外光谱具有药物和蛋白的叠加特性,其最大吸收峰在波长277 nm处,说明载体蛋白BSA与DIF成功偶联,可用于动物免疫。检测抗原DIF-OVA的最大吸收峰在波长276 nm处,说明载体蛋白OVA与DIF成功偶联,可用作检测抗原进行包被。间接ELISA检测结果表明所得小鼠抗血清效价较高,均达1∶32 000以上,说明有特异性抗体产生。[结论]成功制备了抗二氟沙星抗体,也进一步证明了药物偶联成功。

[ Objective ] The research aimed to difloxacin（DIF） antibody. [ Method ] Using N-hydroxysuccinimide active ester （NHS） meth- od, difloxacin was coupled with bovine serum albumin（BSA） and ovalbumin（OVA） to prepare the immunizing antigen DIF-BSA and detecting antigen DIF-OVA. Mice were immunized by using DIF-BSA to obtain the antibody with high titer. [ Result] The ultraviolet spectrum of immu- nizing antigen DIF-BSA had the stacking characteristics of medicament and protein and its. maximum absorption peak appeared at the wave- length of 277 nm, which indicated that vector protein BSA was successfully coupled with DIF and it could be used in animal immunization. The maximum absorption peak of detecting antigen DIF-OVA appeared at the wavelength of 276 nm, indicating vector protein OVA was suc- cessfully coupled with DIF and it could be taken as the detecting antigen for coating. The results of indirect ELISA detection showed that the antiserum titers of mice were higher and over 1：32 000, which indicated that the specific antibody was produced. [Conclusion] The polyclonal antibody a^ainst DIF was successfully prepared, which further proved that I）IF were successfully coupled.