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目的首次合成并鉴定马钱苷(loganin)人工抗原,为进一步制备马钱苷单克隆抗体,建立相关免疫分析方法奠定基础。方法采用高碘酸钠氧化法合成马钱苷免疫原(loganin-BSA)和包被原(loganin-OVA);应用基质辅助激光解析电离-飞行时间质谱(MALDI-TOF-MS)法鉴定马钱苷人工抗原是否偶联成功;通过酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)方法检测免疫小鼠血清中抗马钱苷抗体效价及其特异性。结果经MALDI-TOF-MS检测,马钱苷与BSA偶联成功。竞争抑制曲线表明,血清中的抗体可与马钱苷产生特异性结合,血清抗体效价在1∶40 000。结论成功合成了马钱苷人工抗原,为马钱苷单克隆抗体的制备及其在中药质量评价和动物体内药动学方面的应用奠定了基础。
OBJECTIVE To synthesize and identify loganin artificial antigen for the first time and lay the foundation for the further preparation of loganin monoclonal antibody and establishment of relevant immunoassay methods. Methods Loganin-BSA and loganin-OVA were synthesized by sodium periodate oxidation method. MALDI-TOF-MS was used to identify the allergens Glycosides artificial antigens were successfully coupled; anti-mouse anti-logarithmic antibody titer and specificity was detected by enzyme-linked immunosorbent assay (ELISA). Results The results of MALDI-TOF-MS showed that the loganin was conjugated with BSA successfully. Competitive inhibition curves indicate that antibodies in serum can specifically bind to loganin with a serum antibody titer of 1: 40,000. Conclusion The stilbin glycoside artificial antigen was successfully synthesized, which provided the basis for the preparation of the loganin monoclonal antibody and its application in the quality evaluation of Chinese medicine and pharmacokinetics in animals.