芦荟苷人工抗原的合成与鉴定

Synthesis and identification of artificial antigen of aloin

目的:合成并鉴定芦荟苷人工抗原,为进一步生产芦荟苷单克隆抗体和研究其药理药效奠定基础。方法:采用高碘酸钠氧化法分别制备芦荟苷免疫原(Alo-BSA)和包被原(Alo-OVA)。用薄层色谱法和紫外光谱法鉴定抗原的合成是否成功;用动物免疫的方法测定其抗原性,用间接ELISA方法检测血清效价;用间接竞争ELISA法检测抗体特异性。结果:经薄层色谱法和紫外光谱法检测,芦荟苷与BSA和OVA均成功偶联;用免疫原Alo-BSA可成功使BALB/c小鼠产生芦荟苷抗体;建立的间接ELISA法测得血清效价为1∶32 000,建立的间接竞争ELISA法证明所得的抗体能特异性地与芦荟苷结合。结论:高碘酸钠氧化法成功合成了芦荟苷的人工抗原,具有良好的免疫原性,能诱发BALB/c小鼠产生针对芦荟苷的特异性抗体,可用于制备芦荟苷单克隆抗体及建立相应免疫分析法。

Objective：To synthesize and identify the aloin artificial antigen,and thus to lay the foundation for the further production of aloin monoclonal antibody and the studies of its pharmacological effect.Methods：Oxidation method using sodium iodide was used to synthesize immunogenic antigen（Alo-BSA）and coating antigen（Alo-OVA）.The ultraviolet spectroscopy and thin layer chromatography were used to identify whether the synthetic antigen was successful.The animal immune method was used for determining the antigenicity,the indirect ELISA method was used to detect serum titer,and the indirect competitive ELISA method was used to detect the antibody specificity.Results：Thin layer chromatography and ultraviolet spectroscopy detection showed aloin was successfully coupled with BSA and OVA.An immunogenic Alo-BSA can successfully make the BALb/c mice produce aloin antibody.The serum titer was measured as 1∶32 000 by the established indirect ELISA method.The established indirect competitive ELISA method proved that the obtained antibody can be specifically combined with aloin.Conclusion：The oxidation method using sodium iodide can be applied in synthestizing artificial aloin antigen which has good immunogenicity and can induce BALB/c mice specifically to produce anti-aloin antibodies.It can be used for the preparation of aloin monoclonal antibody and establishment of corresponding immune analytical methods.