Phrompittayarat Watoo, Putalun Waraporn, Tanaka Hiroyuki, Jetiyanon Kanchalee, Wittaya-Areekul Sakchai, Ingkaninan Kornkanok
Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences, Naresuan University, Phitsanulok 65000, Thailand.
Phytochem Anal. 2007 Sep-Oct;18(5):411-8. doi: 10.1002/pca.996.
Bacopa monnieri (L.) Wettst. (Brahmi) is an Ayurvedic medicinal plant used for centuries as a memory enhancer. Dammarane-type triterpenoid saponins classified as pseudojujubogenin and jujubogenin glycosides are reported as the active components in this plant. In this study, the monoclonal antibody (MAb) against bacopaside I, a major pseudojujubogenin glycoside found in Brahmi, was produced and characterised. Bacopaside I was conjugated with bovine albumin serum to prepare an immunogen. Hybridomas secreting a monoclonal antibody against bacopaside I was produced by fusing splenocytes from an immunised mouse and SP2/0-Ag14 myeloma cells. The results showed that the antibodies were raised specifically against pseudojujubonin glycosides. An enzyme-linked immunosorbant assay (ELISA) using anti-bacopaside I MAb was performed in the range of 1.95-62.5 ng/mL of bacopaside I, with a limit of detection of 0.5 ng/mL. The method was validated and the practical use of the ELISA for analysing saponin glycosides from Brahmi was demonstrated.
假马齿苋（Bacopa monnieri (L.) Wettst.，即“Brahmi”）是一种阿育吠陀药用植物，数百年来一直被用作记忆增强剂。据报道，该植物中的活性成分是归类为假酸枣皂苷元和酸枣皂苷元糖苷的达玛烷型三萜皂苷。在本研究中，制备并表征了针对假马齿苋中主要的假酸枣皂苷元糖苷——假马齿苋皂苷I的单克隆抗体（MAb）。将假马齿苋皂苷I与牛血清白蛋白偶联以制备免疫原。通过将免疫小鼠的脾细胞与SP2/0-Ag14骨髓瘤细胞融合，产生了分泌针对假马齿苋皂苷I的单克隆抗体的杂交瘤。结果表明，这些抗体是针对假酸枣皂苷元糖苷特异性产生的。使用抗假马齿苋皂苷I单克隆抗体进行的酶联免疫吸附测定（ELISA）在假马齿苋皂苷I浓度为1.95 - 62.5 ng/mL范围内进行，检测限为0.5 ng/mL。该方法经过验证，并证明了ELISA在分析假马齿苋中皂苷糖苷方面的实际应用。
