A time-resolved fluoroimmunoassay for 18-oxocortisol and 18-hydroxycortisol. Development of a monoclonal antibody to 18-oxocortisol

Patients with primary aldosteronism and with glucocorticoid-suppressible aldosteronism excrete in the urine excessive amounts of the hybrid steroids 18-hydroxycortisol and 18-oxocortisol. The measurement of these steroids aids in the differential diagnosis of various adrenal disorders. We have produced mouse monoclonal antibodies against 18-oxocortisol and polyclonal antibodies against 18-hydroxycortisol and describe a time-resolved fluoroimmunoassay (TR-FIA) technique for the measurement of these steroids in the urine. We have also compared this assay with an ELISA technique for these compounds. We also describe the preparation of in-house Eu(III)-labeled avidin and an enhancement solution and compared to a commercially available Eu(III)-labeled streptavidin and enhancement solutions. The monoclonal antibodies against 18-oxocortisol are sensitive and have a high level of specificity. The TR-FIA technique using in-house prepared reagents or commercial ones were indistinguishable from each other, but at a significant saving. The TR-FIA technique was more sensitive and had a greater precision than the ELISA technique for both steroids.