Abstract: Soy glycinin is a very important allergen in soy protein. In this experiment, glycinin was purified from soy protein by alkali-dissolved acid precipitation method, and it was purified through CL-6B gel column chromatography to obtain a higher purity. The sample protein was dialyzed to remove salt, and then its concentration was determined. Finally, the protein purity was analyzed by SDS-PAGE electrophoresis. The test results showed that the purity of soybean glycinin was higher than 80%, which meets the requirements of the follow-up experiments. Then the purified glycinin was used as an immunogen, and mixed with adjuvant to be fully emulsified. Six Balb/c female mice (7-8 weeks old) were immunized with it by subcutaneous injections at multiple points. Three weeks after the first immunization, the second immunization was performed, followed by immunization every two weeks. One week after the fourth immunization, the serum was obtained by cutting the tail of the mouse, then the serum was tested for its titer and related immunological characteristics. After the test, the titers of the polyclonal antiserum of 4 mice were above 1:6 400, which met the experimental requirements. Among them, the titers of mice 1 and 2 were the highest (1:12 800). The dilution multiple of the OD(450) closest to 1 in the titer determination was selected as the polyclonal antiserum dilution of the sensitivity determination, and the soy glycinin solution was used as the inhibitor to carry out the sensitivity detection experiment, and finally the respective IC₅₀ values were calculated. The sensitivity test results showed that the sensitivity of the four mice was 602 ng/mL, 3 802 ng/mL, 630 ng/mL, and 1 584 ng/mL. Among them, mouse No.1 had the strongest polyclonal antiserum sensitivity. The polyclonal antiserum of mouse No.1 which has the highest titer and strongest sensitivity was selected for specific identification. No reaction was found between the serum/polyclonal antiserum and wheat/sesame protein. Although it has a slight cross-reaction with peanut protein and β-conglycinin, the cross-reaction rate was below 5%. Hence, the polyclonal antiserum had a strong specificity. Subsequently, the polyclonal antiserum of mouse No.1 was used in western blotting experiments. It was found that the polyclonal antiserum reacted strongly with the A2 subunit of glycinin rather than other subunits. It may be caused by the high content of A2 subunit in soybean glycinin and its strong allergenicity. This result again proved that the mouse-derived polyclonal antiserum had a very strong specificity. In this experiment, we obtained a mouse-derived polyclonal antiserum with a high titer, strong sensitivity and excellent specificity. This study also provides a theoretical basis for the development of rapid immunological detection methods and the guidelines for the desensitization of glycinin.