甘草酸单克隆抗体的制备及竞争ELISA法的建立

Preparation of monoclonal antibody against glycyrrhizic acid and development of competitive ELISA

目前针对甘草酸的检测方法存在成本高、效率低的缺点,本研究通过碳二亚胺法合成甘草酸免疫原并制备出高效价高灵敏度的单克隆抗体,以建立一种基于间接竞争法的酶联免疫检测技术。本研究对常见甘草类型进行甘草酸含量测定并用HPLC法验证。结果表明:抗体效价>5×104,纯化抗体的蛋白浓度为10.03 mg/m L。竞争ELISA法的最小检测极限为1.62 ng/m L,检测范围为1.62-200 ng/m L。ELISA方法测定的数据与HPLC方法测定的数据有很好的相关性。该方法具有简单快速,效率高,成本低的特点,对甘草酸的含量测定有重要意义。

At present, the detecting methods for glycyrrhizic acid has low sensitivity and high cost. In this study, the glycyrrhizic acid immunogen was synthesised through the carbodiimide（CDI） method. The high sensitivity monoclonal antibody was prepared to establish indirect competition law- based enzyme- linked immune detection technology. The results showed that the antibody titer was above 1∶50000, and the concentration of purified antibody was 10.03 mg/m L. Moreover, the minimum detection limit of the competitive ELISA was 1.62 ng/m L, with detection range between 1.62 ng/m L and 200 ng/ml. The analysis result of the ELISA method was highly consistent with that of HPLC. The established competitive ELISA has some advantages, such as simplicity,high efficiency and low cost, which lays a foundation for rapid detection of glycyrrhizic acid in samples.