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比较小分子药物磺胺二甲基嘧啶(SM2)用两种不同方法合成完全抗原的免疫效果,通过比较这两种不同方法合成的完全抗原的免疫效果,为进一步的SM2单克隆抗体的制备和SM2残留检测奠定基础。分别采用重氮化(直接连接)、戊二醛法(引入间隔臂)将药物磺胺二甲基嘧啶(SM2)偶联到牛血清白蛋白(BSA)上,并用基质辅助激光解析电离飞行时间质谱鉴定完全抗原合成,确定其偶联比,然后用合成的完全抗原免疫小鼠,免疫周期结束后眼眶取血,用酶联免疫分析法测定血清效价,比较两种不同方法合成的完全抗原的免疫效果。通过基质辅助激光解析电离飞行时间质谱鉴定,重氮化法和戊二醛法都成功合成了磺胺二甲基嘧啶(SM2-BSA)完全抗原,偶联比分别为13.6∶1和12.6∶1。通过小鼠血清的酶联免疫分析法检测,进一步证明实验成功合成了SM2完全抗原,并能有效的刺激机体产生良好的免疫反应。
Compared with the small molecule drug sulfamethazine (SM2), the immune effect of the complete antigen was synthesized by two different methods. By comparing the immune effects of the complete antigen synthesized by the two different methods, further SM2 monoclonal antibody preparation and SM2 Residue testing laid the foundation. The sulfadimethoxine (SM2) was conjugated to bovine serum albumin (BSA) by diazotization (direct attachment) and glutaraldehyde method (introduction of spacer), respectively. The matrix-assisted laser desorption ionization time of flight mass spectrometry The synthesis of complete antigen was identified and its coupling ratio was determined. The mice were then immunized with the synthetic complete antigen. After the end of the immunization period, orbital blood was taken and the serum titer was determined by enzyme-linked immunosorbent assay. Immune effect. Sulfadimethylpyrimidine (SM2-BSA) complete antigen was successfully synthesized by matrix-assisted laser desorption ionization time of flight mass spectrometry. The coupling ratios were 13.6:1 and 12.6:1, respectively. By the mouse serum enzyme-linked immunosorbent assay to further prove that the experiment successfully synthesized SM2 complete antigen, and can effectively stimulate the body to produce a good immune response.