3-氨基-2-嗯唑烷酮半抗原、人工抗原的合成及鉴定

Synthesis and characterization of AOZ hapten and artificial antigens

以2,4-二硝基氟苯为衍生剂,建立了3-氨基-2-噁唑烷酮(AOZ)的高效衍生方法,成功获得了衍生物F-AOZ,并进一步通过硝基的选择性还原合成了新的半抗原AM-AOZ,经电喷雾电离质谱(ESI-MS)和核磁共振(NMR)鉴定F-AOZ与AM-AOZ合成成功;采用重氮化法和戊二醛交联法将半抗原AM-AOZ分别与牛血清蛋白BSA和卵清白蛋白OVA进行偶联,合成了AOZ免疫原和包被原,经紫外吸收法进行成功鉴定;同时,分别采用紫外扫描法和三硝基苯磺酸法(TNBS)测定偶联比。另外,采用分子模拟手段对抗原表面半抗原决定簇的前线分子轨道分布情况进行了模拟分析,讨论了前线轨道的分布对可能获得的抗体特性的影响,为进一步制备高质量抗体提供理论支持。

A new derivant of 3 - aminooxazolidin-2 - oxazolidinone （AOZ） （ F- AOZ） was synthesized by using 1-fluoro-2,4-dinitrobenzene as material with high efficiency. The hapten against AOZ （AM-AOZ）was further synthesized by introducing a spacer through the nitro group of F-AQZ.The results of ESI-MS and NMR indicated that F-AOZ and AM-AOZ were both synthesized successfully.Hapten AM-AOZ was conjugated to BSA and OVA by using diazotization and glutaraldehyde methods respectively to obtain the artificial antigens.The antigens were characterized by UV spectrum, which indicated the successful synthesis of antigens. The ratios of the hapten conjugated to protein were determined by UV spectrum and 2,,4,6-trinitrobenzene sulfenicacid method （TNBS）. Frontier molecular orbital （FMO）distribution of conjugate mimics was analyzed by molecular modeling. The research laid a foundation for further discovering the structure-activity relations （SAR） of small molecule hapten and developing antibody and immunoassay method.