Development of chemiluminescence monitored immunoassays for steroid hormones

Solid-phase chemiluminescence based immunoassay methods for the measurement of steroids and urinary steroid conjugates are described. The assays incorporate the following features: (i) a steroid-chemiluminescent marker conjugate serves as the labeled ligand; (ii) polyclonal or monoclonal antibodies covalently coupled to polymer beads are utilized as the specific immunoadsorbent; and (iii) a centrifugation step after the binding reaction is used to remove interferring luminescent compounds along with unbound steroid. These assays proved satisfactory with respect to sensitivity, precision and accuracy and offer significant advantages over existing methods in speed, safety and economy. Moreover, the use of monoclonal antibodies provides optimal specifity and rigorous standardization.