牡荆素人工抗原的合成与免疫原性的鉴定

Synthesis and identification of artificial antigens of vitexin

目的合成牡荆素的人工抗原,并对其免疫原性进行了鉴定。方法利用高碘酸钠氧化法偶联牡荆素制备牡荆素-牛血清白蛋白的免疫抗原和牡荆素-卵清蛋白的包被原;用紫外分光光度法和薄层色谱法鉴定小分子与载体蛋白偶联反应,并用间接ELISA测定免疫抗原免疫小鼠抗体效价,间接竞争ELISA（ic-ELISA）检测抗体特异性。结果紫外扫描和薄层色谱结果表明牡荆素与牛血清白蛋白/卵清蛋白偶联成功,小鼠经牡荆素-牛血清白蛋白免疫后,体内产生了抗牡荆素抗体,效价在1∶16 000以上,线性检测范围为0.028 8-18μg/m L。结论合成的牡荆素人工抗原和专属酶联免疫分析法可用于后续的单克隆抗体的制备。

AIM To synthesize the artificial antigen of vitexin and identify its immunogenicity. METHODS Carrier protein was employed to couple vitexin to form immunogenic antigen（ vitexin-BSA） and coating antigen（ vitexin-ovalbumin） via sodium iodide oxidation method. UV spectrometry and thin layer chromatography（ TLC）method was used in the characterization of the synthesis. Balb / c mice were immunized with the prepared vitexinBSA immunogenic antigen. Their tilter and specificity of the anti-serum was examined and detected by indirect ELISA and indirect competitive ELISA（ ic-ELISA）,respectively. RESULTS UV spectrometry and TLC conformed that vitexin conjugated with ovalbumin and BSA were carried out. Receiving vitexin-BSA,the mice produced anti-vitexin antibodies specifically,whose titer was above 1 ∶ 16 000,and their linear range detection was from 0. 028 8 μg / m L to 18 μg / m L. CONCLUSION The synthesized artificial antigen of vitexin and special ELISA can be applied to the preparation of monoclonal antibodies.