Preparation of Anti-Glycyrrhetinic Acid Monoclonal Antibody for Application in an Indirect Competitive Enzyme-Linked Immunosorbent Assay

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  • Shunsuke Fujii
    Department of Health and Nutrition, Faculty of Health Management, Nagasaki International University, Sasebo, Japan
  • Takuhiro Uto
    Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Japan
  • Shuichi Nomura
    Department of Health and Nutrition, Faculty of Health Management, Nagasaki International University, Sasebo, Japan
  • Yukihiro Shoyama
    Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Japan

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ABSTRACTGlycyrrhetinic acid is a major metabolite of glycyrrhizin, which is one of the main components of licorice roots and is considered to be one of the pharmacologically active substances in licorice. A new hybridoma cell line, named G-2A6, was generated by fusing mouse myeloma cells and splenocytes, which were immunized using glycyrrhetinic acid-keyhole limpet hemocyanin to produce a monoclonal antibody (mAb) against glycyrrhetinic acid. By using the anti-glycyrrhetinic acid mAb, we attempted to develop a simple, rapid, and highly sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA). The developed icELISA had a range from 3.91 to 125 ng/ mL with low coefficients of variation (less than 5%) and demonstrated a high recovery rate of glycyrrhetinic acid spiked into licorice powder (average = 101.76%). In addition, the icELISA could determine the glycyrrhetinic acid concentration in glycyrrhetinic acid-spiked human serum with simple pretreatment, which suggests that the developed ELI...

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