COMPETITIVE IMMUNOASSAY FOR HAPTEN USING ENZYME LABELLED ANTIBODY

摘要

IN THE UNITED STATES PATENT OFFICEAbstract-of the Disclosure:According to the present invention there is provided asensitive quantitative assay for the determination of a hapten "X"which comprises: preparing a conjugate of the hapten X which conjugateis adapted to ellicit antibody formation, and injecting a mammal withsaid conjugate, resulting in a specific antibody against the saidhapten, "anti-X"; preparing a conjugate of the hapten X and of anotherentity Y, which is either another hapten or a larger molecule,injecting a mammal with the entity Y if it is a larger molecule,or with a conjugate thereof if it is a small molecule which doesnot by itself ellicit antibody formation, so as to form anti-Yantibodies, preparing a conjugate of a suitable enzyme and theantibody against Y, i.e. an (enzyme)-(anti-Y)-conjugate; adsorbingthe antibodies against X, i.e. anti-X on a solid support, conitactingsaid solid support with a mixture containing the unknown quantity ofhapten X and a known quantity of the X-Y conjugate; removingunreacted X and X-Y, leaving all anti-X sites occupied, addingenzyme-labelled anti-Y antibody; adding a suitable substrate resultingin a color reacting and measuring the intensity of color indicatingthe quantity of bound enzyme and deducing from calibration curvesthe quantity of the hapten X; and to a kit for carrying out suchassay.