梓醇人工抗原的合成、鉴定及免疫原性的初步研究

Synthesis,identification of artificial antigen of catalpol and preliminary study of immunogenicity

基于单克隆抗体的免疫分析方法具有高特异性、高灵敏性等特点,在中药的质量控制和动物体内微量成分的检测等方面均具有重要的意义,该方法建立的前提条件就是成功制备中药活性小分子化合物的人工抗原。该研究采用高碘酸钠氧化法,首次制备了梓醇的人工抗原,通过基质辅助激光解析电离-飞行时间质谱法(MALDI-TOF-MS)和分子排阻色潜法鉴定梓醇与牛血清白蛋白(BSA)偶联成功,用间接ELISA方法检测免疫小鼠血清中抗梓醇抗体效价在1∶8 000以上,成功制备梓醇的人工抗原,为下一步单克隆抗体的制备和相关免疫方法的建立奠定基础。

The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine （TCM） and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry （MALDI-TOF-MS） and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1：8 000. The artificial antigen of catalpol was successfully synthesized.