| 摘要: |
| 【目的】合成并鉴定斑蝥素人工抗原，为建立斑蝥素的酶联免疫分析方法提供依据。【方法】对斑蝥素进行化学修饰，引入羟基合成N 羟乙基斑蝥酰亚胺（BY），并经MS、NMR、IR法对其结构进行鉴定。进一步经琥珀酸酐法合成琥珀酸酯（BYS）并纯化，在此基础上，分别采用混合酸酐法和碳二亚胺法将半抗原与载体蛋白偶联，合成斑蝥素的人工抗原（BYS-HS-BSA、BYS-TY-BSA、BYS-HS-OVA、BYS-TY-OVA），并经MS、IR法对人工抗原进行定性鉴定。免疫原免疫兔子获得抗斑蝥素多克隆抗体并测定其效价。【结果】成功合成了一种斑蝥素半抗原，与载体蛋白BSA、OVA偶联后得到免疫原（BYS-HS-BSA、BYS-TY-BSA）和包被原（BYS-HS-OVA、BYS-TY-OVA），免疫新西兰大白兔获得多克隆抗体，效价分别为8 000和16 000。【结论】初步合成并鉴定了斑蝥素人工抗原，在设计合成斑蝥素的半抗原时不能破坏其关键的环结构。 |
| 关键词: 斑蝥素 半抗原 人工抗原 多克隆抗体 |
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| 基金项目:公益性行业（农业）科研专项（200903052）；陕西省13115科技创新工程重大科技专项（2007ZDKG-14）；陕西省科学技术研究发展计划项目（农业攻关）（2007K01-17） |
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| Synthesis and identification of artificial antigen for Cantharidin |
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| Abstract: |
| 【Objective】This research aimed to synthesize and identify artificial antigen of Cantharidin to establish an indirect competitive enzyme-linked immunosorbent assay for further research.【Method】To this end,Cantharidin was modified to the hapten,N-ethoxylcantharidimide(BY).The hapten was characterized by MS,NMR and IR.Then synthsis of BYS was done by mixed anhydride method.After that,to get the immunogens and coating antigens,the hapten was coupled to carrier proteins BSA and OVA by means of mixed anhydride reaction and Carbodiimide method respectively.These four artificial antigens,namely BYS-HS-BSA,BYS-TY-BSA,BYS-HS-OVA,BYS-TY-OVA,were characterized and verified by IR and MS. Both immunogens,BYS-HS-BSA and BYS-TY-BSA,were used to immunize rabbits to produce polyclonal antibodies against Cantharidin and to measure the titer.【Result】Hapten of Cantharidin was synthesized successfully,and conjugated with BSA and OVA to form immunogens (BYS-HS-BSA,BYS-TY-BSA) and coating antigens (BYS-HS-OVA,BYS-TY-OVA).Two types of anti-cantharidin polyclonal antibodies (BYS-HS-Ab,BYS-TY-Ab) with titers (8 000 and 16 000) were obtained.【Conclusion】Artificial antigen of cantharidin was preliminarily synthesized and indentified.Besides,when synthesizing the hapten,the ring structure of cantharidin,which is a key one,should not be breached. |
| Key words: Cantharidin hapten artificial antigen polyclonal antibody |
