Preparation and identification of monoclonal antibodies against glycyrrhizic acid

Objective:To prepare a specific monoclonal antibody against glycyrrhizic acid(GA).Methods:BALB/c mice were immunized with the prepared GA-BSA conjugate.When the serum detection was positive,spleen cells from the mice were isolated and fused with SP2/0 myeloma cells at a ratio of 10∶ 1.Monoclonal hybridoma cells were screened by indirect ELISA and limited dilution.The ascites antibodies were induced and prepared by the positive cell line,and then were purified by caprylic acid/ammonium sulfate precipitation and tested by cross reaction.Results:The monoclonal antibody hybridoma cell line GA-Mab-DF5 steadily secreting glycyrrhizic acid antibodies was obtained,the purity of ascites antibodies was up to 98%,and the recovery rate was 80%.The linear range was 4 μg·mL-1 to 64 μg·mL-1(R2=0.9986)and no cross-reactivity was detected with BSA,gelatin,cholic acid,deoxycholic acid,paeoniflorin,baicalin and so on.Conclusion:The anti-GA antibody-secreting hybridomas are obtained with high sensitivity and specificity,which lays a foundation for the trace detection and immunoaffinity column preparation of glycyrrhizic acid in samples.