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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 monoclonal antibody-based immunoassay method is of high specificity and high sensitivity. It has important significance in the quality control of Chinese medicine and the detection of trace components in animals. The precondition of this method is the successful preparation of traditional Chinese medicine Artificial antigen for small molecule compounds. In this study, the artificial antigen of catalpol was prepared for the first time by the sodium periodate oxidation method. The catalysis of catalpol with bovine serum was determined by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) Albumin (BSA) conjugate was successfully used to detect anti-catalpol antibody titer in the serum of the immunized mice by 1: 8000 or more by indirect ELISA. The artificial antigen of catalpol was successfully prepared for the next monoclonal antibody preparation and related The establishment of immune methods laid the foundation.