Objective To construct artificial immunogen and prepare specific antiserum of diammonium glycyrrhizinate (DG). Met- hod Immunogen DG-BSA and coating antigen DG-OVA were prepared by carbodiimide method. The polyclonal antiserum was produced from two big ear rabbits immunized with conjugates DG-BSA, and then the antiserum was determined with indirect ELISA method. Results The ideal coating antigen concentration was 200 ng/mL and the available antiserum dilution was 1∶10 000. The titers of antiserum were 1/25 600 for male rabbit and 1/51 200 for female one. Conclusion The artificial immunogen could be synthesized by carbodiimide method and the specificity of the polyclonal anti-DG antibodies could be produced. The methods lay a foundation for determining DG fast and effectively in therapeutic drug monitoring (TDM).