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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 artificial antigens of aloe glycosides, and to lay the foundation for the further production of monoclonal antibodies against aloin and its pharmacological effects. Methods: Alo-BSA and Alo-OVA were prepared by sodium periodate oxidation. The antigenicity was determined by TLC and UV spectrophotometry. The antigenicity was determined by animal immunization. The serum titer was measured by indirect ELISA and the antibody specificity by indirect competitive ELISA. Results: Aloin was successfully coupled with BSA and OVA by TLC and UV spectrophotometry. The Alo-BSA immunogen was successfully used to produce albino antibody in BALB / c mice. The indirect ELISA The serum titer was 1:32 000. The established indirect competitive ELISA demonstrated that the obtained antibody could specifically bind to aloin. CONCLUSION: The artificial antigen of aloin glycoside is synthesized successfully by sodium periodate oxidation method and has good immunogenicity. It can induce specific antibody against aloe glycosides in BALB / c mice and can be used to prepare albino glycoside monoclonal antibody. Corresponding immunoassay.