| 摘要: |
| 【目的】合成并鉴定天名精内酯酮人工抗原，制备天名精内酯酮多克隆抗体，为天名精内酯酮作用靶标的免疫化学定位奠定基础。【方法】通过天名精内酯酮4位羰基和盐酸氨基脲氨基的亲核加成反应制备半抗原天名精内酯酮缩氨基脲（CN），使用MS和NMR对其结构进行鉴定；采用戊二醛法将半抗原和载体蛋白牛血清蛋白（BSA）和卵清蛋白（OVA）偶联，合成天名精内酯酮免疫原（CN-BSA）和包被原（CN-OVA），采用紫外吸收扫描法、红外光谱法对免疫原及包被原进行鉴定；用制备好的免疫原免疫健康BALB/c小鼠获得天名精内酯酮多克隆抗体，通过间接非竞争ELISA法测定其效价。【结果】成功制备了天名精内酯酮免疫原（CN-BSA）和包被原（CN-OVA），获得了效价为128 000的天名精内酯酮多克隆抗体。【结论】成功制备了天名精内酯酮多克隆抗体。 |
| 关键词: 天名精内酯酮 半抗原 人工抗原 酶联免疫吸附分析 |
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| 基金项目:国家自然科学基金项目（31272074）；陕西省自然科学研究计划项目（2012JQ3004）；中央高校基本科研业务费专项(ZD2012004) |
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| Preparation of polyclonal antibody against carabrone |
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HE Zeyu,HAN Lirong,LIU Xueru,et al
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| Abstract: |
| 【Objective】This study synthesized and identified artificial antigen of carabrone,and prepared polyclonal-antibody.【Method】 Reaction of carabrone and semicarbazide hydrochloride was conducted to obtain hapten carabrone semicarbazone,which was characterized by MS and NMR spectrum.Then,the haptens were conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) for generation of immunogen (CN-BSA) and coating antigen (CN-OVA),respectively.The structures of immunogen and coating antigen were identified by UV and IR spectrum.BALB/c mice were immunized with CN-BSA and polyclonal antisera were produced.The titer of antiserum was also detected by indirect non-competitive enzyme-linked immunosorbent assay.【Result】Hapten was synthesized,and immunogen (CN-BSA) and coating antigen (CN-OVA) were obtained successfully.Polyclonal-antibody against carabrone was prepared and the titer of antiserum was 128 000.【Conclusion】The artificial antigen and polyclonal-antibody against carabrone was prepared successfully. |
| Key words: carabrone hapten artificial antigen ELISA |
