桔梗皂苷D人工抗原的合成及鉴定

目的:合成桔梗皂苷D(PlatycodinD,PD)的人工完全抗原,为制备桔梗皂苷D的单克隆抗体及建立相应的免疫学分析方法奠定基础。方法:采用高碘酸钠(NaIO4)氧化法分别合成桔梗皂苷D免疫抗原(PD-BSA)和包被抗原(PD-OVA)。用薄层色谱法鉴定人工抗原是否偶联成功;用动物免疫法测定其抗原性;并融合后筛选阳性孔。结果:薄层色谱法显示PD与BSA/OVA偶联成功;用合成的PD-BSA免疫小鼠后可使小鼠体内产生抗PD抗体,效价在1∶8000以上;融合可得出阳性孔。结论:成功合成了PD人工免疫原和包被原,可用于下一步PD单克隆抗体的制备及建立相应免疫学分析方法。

Synthesis and Ldentification of Artificial Antigen of Platycodin D

Objective ： To establish an y for analyzing platycodin D （PD） and to synthesize the artificial antigen platycodin D - bovine serum albumin （ PD - BSA） and the artificial coated antigen platycodin D - ovalbumin （ PD - OVA）. Methods：PD was conjugated with BSA and OVA respectively by sodium periodate oxidation method. The characterization of the synthesis was examined by thin layer chromatography （TLC）. The titer and specificity of antibody in serum from Balb/c mice was detected by indirect competitive enzyme - linked immunosorbent assay （ IC - ELISA）. When the serum detection was positive, spleen cells from the mice were isolated and fused with SP2/0 myeloma cells, and then the cells were also screened by indirect ELISA. Results ： According to the detection of TLC, the PD was successfully conjugated with BSA and OVA. The test result showed the anti - PD antibody obtained from immunized mice could bind to PD specially and the titer up to 8000. Several positive wells were gained. Conclusion ： The artificial immunogen PD - BSA and coated antigen PD - OVA were successfully synthesized, which can be used further for preparing monoclonal antibody.