MONOCLONAL ANTIBODIES FOR DIRECT STEROID IMMUNOASSAY

THE GOAL FOR PHASE I IS TO OBTAIN A HIGHLY SPECIFIC MONOCLONAL ANTIBODY (MAB) FOR TESTOSTERONE WITH WHICH AN IMMUNOMETRIC ASSAY FOR TESTOSTERONE WILL BE DEVELOPED THAT HAS SUPERIOR ANALYTICAL ACCURACY THAN COMMERCIAL KITS CURRENTLY AVAILABLE. THE KEY ASPECT IS TO OBTAIN A MAB WITH THE NECESSARY SPECIFICITY AND AFFINITY. IMMUNOASSAYS CURRENTLY AVAILABLE SUFFER FROM OVERESTIMATION OF THE STEROID CONCENTRATION IN SERUM, PROBABLY DUE TO AN INSUFFICIENT SPECIFICITY OF THE ANTIBODY. A 15ALPHA-CARBOXYMETHYL DERIVATIVE OF TESTOSTERONE COUPLED TO KLH USED AS AN ANTIGEN SHOULD ELICIT A MAB WITH THE REQUISITE SPECIFICITY. THE SPECIFICITY OF THE MAB SELECTED WILL BE DETERMINED BY ASSAYING CROSS-REACTIVITY WITH OTHER STEROIDS AND TESTING WITH SERUM SAMPLES CONTAINING CHEMICALLY DETERMINED AMOUNTS OF STEROID. AFTER SELECTION OF A HIGHLY SPECIFIC MAB, AN IMMUNOMETRIC ASSAY UTILIZING A FLUORESCENT ENDPOINT WILL BE DEVELOPED FOR COMMERCIAL APPLICATION. THIS WORK WILL SERVE AS A MODEL FOR DEVELOPMENT OF A COMPLETE LINE OF ASSAYS FOR CLINICALLY IMPORTANT STEROIDS.