| In order to establish a rapid immunological detection method for the arachin allergens,the arachin was extracted by alkali solution and acid precipitation method, and its purity was detemined by SDS-PAGE, and 5 Balb/c female mice aged 6 to 8 weeks were immunized to prepare the arachin mouse-derived polyclonal antiserum,the polyclonal antiserum immunological characteristics were identified. The results showed that the titers of the polyclonal antiserum obtained after the end of immunization were all above 1∶51 200, and all of them had certain sensitivities, among which the polyclonal antiserum from 2# mouse had the best sensitivities, with a half-inhibitory concentration of 320 ng/mL.The polyclonal antiserum from 2# mouse was not sensitive to conarachin, glycinin, β-conglycinin, bovine serum albumin, gluten protein, and skimmed milk powder,and the cross-reaction rate was low(all below 05%),indicating that the specificity of the polyclonal antiserum prepared was relatively strong. Western blot experiment results indicated the successful preparation of arachin mouse-derived polyclonal antiserum. In conclusion, a arachin mouse-derived polyclonal antibody with high titer, strong sensitivity and excellent specificity is obtained, which provide the material foundation for the preparation of monoclonal antibodies and the research of rapid immunological detection methods. |
