Construction of a monoclonal antibody against glabridin (2G4) and development of an enzyme-linked immunosorbent assay.

Abstract

Glabridin is a unique isoflavonoid found only in Glycyrrhiza glabra L. The pharmacological effects of glabridin are well established, especially for beauty- and wellness-related uses, such as antioxidant, anti-inflammatory, ultraviolet (UV) protection, and skin-lightening effects. Therefore, glabridin is often found in commercial products such as creams, lotions, and dietary supplements. This study aimed to develop an enzyme-linked immunosorbent assay (ELISA) using a glabridin-specific antibody. Immunogen conjugation of glabridin-bovine serum albumin was performed via the Mannich reaction, and the resulting conjugates were injected into BALB/c mice. Subsequently, hybridomas were produced. An ELISA method for glabridin determination was developed and validated. A highly specific antibody against glabridin was produced using clone 2G4. The assay range for the determination of glabridin was 0.28-7.02 μg/ml, with a detection limit of 0.16 μg/ml. The validation parameters in terms of accuracy and precision met the acceptable criteria. Standard curves of glabridin in various matrices were compared to evaluate the matrix effect on human serum using ELISA. Standard curves of the human serum and water matrix were obtained in the same manner, and the measurement range was 0.41-10.57 μg/ml. The developed ELISA method was used to quantify glabridin in plant materials and products with high sensitivity and specificity, and has potential applications in quantifying compounds in plant-derived products and human serum samples.