目的 利用种属特异性蛋白建立一种生药甘草的免疫学鉴定方法。方法 选用生药甘草为实验对象 ,利用抗甘草总抗原血清并结合 Western- blot的结果确定甘草种属特异性蛋白 (RGP)。分离纯化该蛋白抗原并制备和生物素标记其特异性抗体 ,建立夹心式 EL ISA法用于甘草的分析鉴定。结果 该方法灵敏度高、专属性强 ,对不同产地及品种的甘草显示了良好的特异性 ,可以用于生药的鉴定和分析。结论 以甘草自身的特异性蛋白为指标的免疫学分析方法,将为生药甘草的品种鉴定及中药材的质量管理提供一条新的途径。

Object To develop an immunoassay method of species-specific protein for Radix Glycyrrhizae (RG) identification. Methods The crude drug of RG as a model was studied in this paper,the antigen of RG protein (RGP) was screened by Western-blot analysis using anti-RG serum. After species-specific protein,RGP was purified,and rabbit antiserum against RGP was prepared,the rabbit IgG of anti-RGP was isolated and biotinylated,then the sandwich enzyme-linked immuosorbent assay (ELISA) was developed. Results The method was only specific to RG,which was harvested from various areas or species,and showed excellent sensitivity and reproducibility. Conclusion The result suggests that the immunoassay method using specific antigen of RGP as a detection objective be a new potential for the unequivocal identification and quality control of RG.

天津市自然科学重点基金资助项目(003805311)