Preparation and ELISA Identification of Mouse Polyclonal Antiserum for βconglycinin

The acquisition of serum byimmunizing animals with antigens is a routine method for the preparation of antibodies. Since β-conglycinin is one of the most important allergenic proteins in soybean protein,the preparation of related antibodies is the basis for the establishment of rapid immunological detection methods. The protein extracted from soybean was identified by SDS-PAGE electrophoresis. With the emulsification of Rreund'sadjuvant,we injected β-conglycinin to Balb/c mice for antiserum preparation. The mice were reimmunizied with β-conglycinin by subcutaneous multiple injection for another four times at each intervals of three weeks. The characteristics of mouse antiserum were determined by enzyme-linked immunosorbent assay(ELISA).The results showed that the immunogen was effective,and the titers of 5 mice were above 1:6 400 after five immunization detected by indirect competitive ELISA,among which antiserum sensitivity of the sixth mouse was the most sensitive,and the of the inhibition concentration (IC₅₀) was 718 ng/mL. The prepared antiserum had no cross-reaction with sesame protein and wheat germ protein,meanwhile the cross-reaction rates of peanut protein and glycinin were less than 10%. This obtainment of mouse antiserum with high sensitivity and specificity may lay the foundation for the rapid immulogical detection of β-conglycinin.