培氟沙星抗体的制备

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培氟沙星抗体的制备
摘要:为了制备培氟沙星(PFLX)抗体,采用N-干手器羟基琥珀酰亚胺活性酯(NHS)法将培氟沙星分别与牛血清白蛋白(BSA)、卵清白蛋白(OVA)过氧化氢含量的测定偶联,电子屏制作制备免疫抗原PFLX-BSA检测抗原PFLX-OVA,PFLX-BSA免疫小鼠以获得高效价的抗体。结果显示,免疫抗原PFLX-BSA紫外光谱具有药物和蛋白的叠加特性,其最大吸收峰在277.5nm,说明载体蛋白BSAPFLX成功偶联,可用于动物免疫。检测抗原PFLX-OVA最大吸收峰在275nm,说明载体蛋白OVAPFLX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樱桃采摘机,均达132 000以上,说明有特异性抗体产生。该研究成功制备了抗培氟沙星抗体,也进一步证明药物偶联成功。
  关键词:培氟沙星旋转滑梯;免疫;多克隆抗体
 
  Production of Multiclone Antibody of PFLX
  Abstract: To produce multiclone antibody of PFLX. PFLX were coupled with bovine serum
albumin(BSA) and ovalbumin(OVA) by using N-hydroxysuccinimide actived ester method to prepare the immune antigen PFLX-BSA and detection antigen PFLX-OVA. The mice were immunized with PFLX-BSA to obtain the polyclonal antibody with high titer. The ultraviolet spectrum of immune antigen PFLX-BSA had the stacking characteristics of medicament and protein and its maximum absorption peak was 277.5 nm,which indicated that vector protein BSA was successfully coupled with PFLX and it could be used in animal immunization.The maximun absorption peak of detection antigen PFLX -OVA was 275nm,indicating vector protein OVA was successfully coupled with PFLX and it could taken as detection antigen for coating. The results of indirect ELISA detection showed that the antiserum titers of mice were higher and reached over 132 000,which indicated that the specific antibody was produced. The polyclonal antibody against PFLX was successfully prepared, which further proved that PFLX were successfully coupled.
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