4 Competitive assay for antibodies

When the interfering substances in the antigen material are not easy to remove, or it is not easy to obtain enough purified antigen, this method can be used to detect specific antibodies. The principle is that the antibody in the specimen competes with a certain amount of enzyme-labeled antibody for binding to the solid-phase antigen. The more antibody in the specimen, the less enzyme-labeled antibody bound to the solid phase, so the positive reaction is lighter than the negative reaction. If the antigen is of high purity, it can be directly coated with the solid phase. If there are interfering substances in the antigen, direct coating is not easy to succeed, you can use the capture coating method, that is, first coat the antibody corresponding to the solid phase antigen, and then add the antigen to form the solid phase antigen. Wash to remove impurities in the antigen, and then add specimens and enzyme-labeled antibodies for competitive binding reaction. There are multiple modes for the detection of antibodies by the competition method. The specimen and the enzyme-labeled antibody can be competitively combined with the solid-phase antigen. This method is generally used for anti-HBc ELISA. Another mode is to add the specimen and the antigen to the solid-phase antibody together for competitive binding. After washing, the enzyme-labeled antibody is added to react with the antigen bound to the solid phase. Anti-HBe detection generally uses this method.
Competition assay
The small molecule antigen or semi-antibody lacks more than two sites that can be used as a sandwich method, so it cannot be measured by the double antibody sandwich method, and the competition method mode can be used. The principle is that the antigen in the specimen competes with a certain amount of enzyme-labeled antigen for binding to the solid-phase antibody. The more the amount of antigen in the specimen, the less the enzyme-labeled antigen bound to the solid phase, and the lighter the final color. This method is often used for ELISA determination of small molecule hormones and drugs.

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