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目的制备磺胺二甲嘧啶(Sulfadimidine,SM2)完全抗原,并进行鉴定。方法采用重氮化偶联法制备SM2免疫抗原(SM2-BSA)和检测抗原(SM2-OVA),定性和定量分析偶联后的完全抗原,并制备SM2单抗。结果经1%琼脂糖凝胶电泳和10%SDS-PAGE分析表明,完全抗原SM2-BSA和SM2-OVA偶联成功;SM2与载体蛋白BSA和OVA的分子结合比分别为23∶1和17∶1;SM2-BSA和SM2-OVA的蛋白浓度分别为3.24和2.29 mg/ml;完全抗原SM2-BSA能刺激小鼠产生高效、特异的抗SM2抗体;SM2单抗与OVA和BSA无交叉反应。结论已成功制备了完全抗原SM2-BSA和SM2-OVA,为下一步建立灵敏度更高、特异性更强、操作更简便的免疫学检测方法奠定了基础。
Objective To prepare and identify the complete antigen of sulfadimidine (SM2). Methods The SM2-BSA and SM2-OVA were prepared by diazotization method. The complete antigen after coupling was qualitatively and quantitatively analyzed and the SM2 mAb was prepared. Results 1% agarose gel electrophoresis and 10% SDS-PAGE analysis showed that the coupling of the complete antigen SM2-BSA and SM2-OVA was successful; the molecular binding ratios of SM2 to BSA and OVA were 23: 1 and 17: 1; the protein concentrations of SM2-BSA and SM2-OVA were 3.24 and 2.29 mg / ml, respectively. The complete antigen SM2-BSA stimulated the mouse to produce efficient and specific anti-SM2 antibody; SM2 mAb did not cross-react with OVA and BSA. Conclusion The complete antigens of SM2-BSA and SM2-OVA have been successfully prepared, which laid the foundation for the next step to establish a more sensitive, specific and easy-to-operate immunological detection method.