摘要
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。
To compare sulfamethazine（ SM2） with complete antigen immune effect of two kinds of synthetic methods,by comparing the immune effect of these two different methods of synthesis of complete antigen,and to lay the foundation for monoclonal antibody SM2 and determination of SM2 residues. Sulfamethazine（ SM2） was conjugated on bovine serum albumin（ BSA） by using diazotization（ direct connection） and glutaraldehyde（ introducing spacer）,and complete antigen synthesis was identified by matrix assisted laser desorption ionization time of flight mass spectrometry（ MALDI-TOF-MS）,the coupling ratio was determined and then mice were immunized with the complete antigen,and blood samples were drawn from orbit after the end of immunizing period,and serum titers were tested by enzyme linked immunosorbent assay（ ELISA）,comparing the immune effection of two methods for the synthesis of complete antigen. The sulfamethazine（ SM2-BSA） antigen was successfully synthesized by diazotization method and glutaraldehyde method with MALDI-TOF-MS,and the coupling ratio were 13. 6∶ 1 and 12. 6∶ 1. By the method of enzyme-linked immunosorbent assay（ ELISA） in mice serum,proving that the SM2 complete antigen was successfully synthesized.
作者
栗世婷
LI Shi-ting(Department of Pharmacy, Inner Mongolia Baotou Steel Hospital, Baotou 014010, China)
出处
《药物生物技术》
CAS
2017年第4期294-298,共5页
Pharmaceutical Biotechnology
关键词
磺胺二甲基嘧啶
完全抗原
合成
鉴定
重氮化法
戊二醛法
Sulfadimidine
Complete antigen
Synthesis
Identification
Diazotization
Glutaraldehyde
Method
作者简介
栗世婷（1980-），女，硕士研究生，主管药师，研究方向：抗生素残留及完全抗原的合成和鉴定，Tel：15540860156，0472-5992956（办），E-mail：337980318@qq．com。
