The invention relates to a
double antibody sandwich
enzyme-linked immuno
sorbent assay (ELISA) method for detecting a peanut allergic component Arah1, which belongs to the technical field of
immunoassay. The method comprises the steps of firstly acquiring peanut
protein with higher purity containing Arah1 from fresh peanut kernels, utilizing the peanut
protein to immunize a mouse to acquire 14 strains of
monoclonal antibodies, and selecting a combination of the
monoclonal antibodies with maximal P / N value as an optimum combination. The
antibody and an
enzyme-labeled
antibody, which are connected onto a
solid-phase carrier, are respectively combined with two antigenic determinants on an
antigen molecule to be detected in a sample to form a
solid antibody-
antigen-
enzyme-labeled antibody immune compound. Since the quantity of the
solid-phase antibody and the enzyme-labeled antibody is excessive relative to the
antigen to be detected in the
reaction system, the formation quantity of the compound is in proportion to the content of the antigen to be detected. The content of the antigen to be detected can be determined by determining the color matter amount (OD value) generated when the enzyme in the compound acts on the added substrate. The
double antibody sandwich
ELISA method is direct and quick, has high sensitivity, high specificity, high accuracy and high precision, is simple to operate and is used for detecting the peanut allergic component Arah1.