摘要
【目的】合成并鉴定苦参碱人工抗原,以获取具有特异性和高亲和性的苦参碱多克隆抗体,为建立苦参碱的酶联免疫分析方法奠定基础。【方法】通过对槐果碱不饱和双键进行亲核加成、叠氮化、催化氢化合成苦参碱半抗原13-氨基苦参碱(ST),并经MS、NMR法对其结构进行鉴定;采用戊二醛法将半抗原与载体蛋白偶联合成苦参碱的人工抗原(ST-BSA和ST-OVA),经紫外光谱扫描法、SDS-PAGE法、红外光谱扫描法对人工抗原进行鉴定;将制备的苦参碱人工抗原免疫健康新西兰大白兔获得苦参碱的多克隆抗体,通过间接非竞争ELISA方法测定其效价。【结果】成功合成了一种苦参碱半抗原13-氨基苦参碱(ST),与载体蛋白BSA、OVA偶联后得到了免疫原(ST-BSA)和包被原(ST-OVA),免疫新西兰大白兔获得多克隆抗体,效价达到128 000。【结论】成功合成了苦参碱人工抗原,为苦参碱免疫分析方法的研究提供了条件。
【Objective】 To establish an indirect competitive enzyme-linked immunosorbent assay for matrine,artifical antigen was synthesized and identified.【Method】 The hapten was designed and synthesized by using sophocarpine through nucleophilic addition,diazotization and hydrogenation then characterized by MS and NMR.The Glutaraldehyde method was used to conjugate hapten to BSA and OVA,the artifical antigen was verified by SDS-PAGE,UV and IR.Polyclonal antisera were isolated from rabbits and the titer was determined by indirect non-competitive ELISA procedure.【Result】 Artificial antigen（ST） was synthesized successfully and conjugated with BSA and OVA to form immunogen（ST-BSA） and coating antigens（ST-OVA）,the titer of anti-matrine serum was 128 000 after immunization of New Zealand white rabbits.【Conclusion】 Results indicated that the polyclonal antibody against matrine would be useful in the further study of IC-ELISA for matrine.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2012年第9期96-102,共7页
Journal of Northwest A&F University(Natural Science Edition)
基金
公益性行业(农业)科研专项"生物源农药创制与技术集成及产业化开发"(200903052)
