A competitive enzyme-linked immunoassay for a soluble analyte in which the analyte in the unknown sample and the same analyte attached to a solid phase (immunosorbent) compete for a selected quantity of a soluble first antibody to the substance; a second or bridging antibody is then linked to any first antibody linked to the immunosorbent as a result of the competitive reaction, following which a soluble enzyme/antibody immune complex is linked to bridging antibody on the immunosorbent. The enzymatic activity associated with the thusly-formed enzyme-immunosorbent complex or with the non-immunosorbent fraction of enzyme is measured to determine the presence and/or concentration of analyte in the unknown sample.
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