Preparation of Antibodies and Preliminary Establishment of Immunoassay Method against Allergen β-Lactoglobulin from Milk
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Abstract:
The β-lactoglobulin antigen was used with injection to immunonize the New Zealand white rabbit and the BALB/c mice so as to get the rabbit polyclonal antibody serum (PcAb) and the monoclonal antibody (McAb), and the titers of antibodies were analyzed; The polyclonal antibody was purified by protein A affinity chromatography and the 3A7 monoclonal antibody was purified by Caprylic acid precipitation. The experiment was operated by the double-antibody sandwich procedure and some samples were detected. The results showed that the titer of polyclonal antibody was over 1:6.56×106, and the polyclonal antibody serum could bind to β-lactoglobulin specifically, and its specificity was low reactivity with casein. The titers of the three strains of monoclonal antibodies were above 106 and the 3A7 monoclonal antibody had almost no cross reactivities with other proteins in milk and other allergen proteins. The accuracy was 100% for the detection of samples.
