Bioluminescent Immunoassay for Testosterone

Radioimmunoassays make up a significant percentage of the immunoassays used in clinicallaboratories. Nonradioactive, immunometric or "sandwich" assays using enzymes for signal generation(EIAs) have generally replaced radioimmunoassays for high molecular weight analytes. Low molecularweight analytes such as the steroid hormones, however, are measured primarily by competitiveimmunoassays using 125I-labeled haptens as the tracer. We manufacture and sell AquaLiteTM, arecombinant form of the calcium-activated photoprotein aequorin, which has been conjugated tomonoclonal antibodies and used as the signal in sensitive bioluminescent immunometric assays for TSHand Interleukin-6. We will synthesize an AquaLite-labeled testosterone for use in a competitiveimmunoassay format for this steroid hormone. The validation of this bioluminescent assay during thefirst phase of this project will define the feasibility for use of AquaLite as a general replacement of 125Iin competitive immunoassays. The assay will be optimized and compared to currently available assays.Once feasibility is established for the testosterone assay, Phase II of this project will be menu expansionby conversion of additional steroid hormone radioimmunoassays to bioluminescent immunoassays andthe development of an FDA approved clinical diagnostic product line for sale by us.