Anti-Ovarian Antibody ELISA

The Anti-Ovarian Ab ELISA is a solid-phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. Microtiter wells are coated with a mix of oocyte proteins. During incubation, anti-oocyte antibodies in the samples (standards, controls, samples) bind to the coated surface of the wells. A wash step removes unbound sample components. Added enzyme conjugate binds to the immobilized antigen-antibody-complexes. The conjugate contains anti-human immunoglobulin antibodies labeled with horseradish peroxidase (HRP). After a wash step to remove all unbound substances, the solid phase is incubated with the substrate solution. The colorimetric reaction is stopped by addition of stop solution, and the optical density (OD) of the resulting yellow product is measured. The intensity of the color is proportional to the concentration of the analyte in the sample. A standard curve is constructed by plotting OD values against concentrations of standards, and concentrations of unknown samples are determined using this standard curve.