HIGH-AFFINITY MONOCLONAL ANTIBODIES FOR ENDOCRINOLOGY

THE LONG-TERM OBJECTIVE OF THIS PROJECT IS TO PROVIDE FOR THE ENDOCRINE COMMUNITY A COLLECTION OF SPECIFIC MONOCLONAL ANTIBODIES (MOABS) WITH ESPECIALLY HIGH AFFINITY FOR A NUMBER OF HORMONES AND RECEPTORS BEING ACTIVELY UTILIZED IN RESEARCH. THE SPECIFIC AIM OF THIS PHASE I PROJECT IS TO DEVELOP ONE OR MORE MOABS WITH THESE CHARACTERISTICS FOR STEROID HORMONES. THE RESULTING MOABS SHOULD BE OF CONSIDERABLE VALUE TO INVESTIGATORS WORKING ON MANY DIFFERENT ENDOCRINE RESEARCH PROBLEMS. EXAMPLES OF USE INCLUDE THE DEVELOPMENT OF MORE SENSITIVE AND SPECIFIC IMMUNOASSAYS, THE DEVELOPMENT OF ANTI-IDIOTYPIC ANTIBODIES USEFUL FOR RECEPTOR ISOLATION, IMMUNO-NEUTRALIZATION OF HORMONAL ACTIVITY, AND IMMUNO-HISTOCHEMICAL LOCALIZATION OF HORMONES. DEVELOPMENT OF DESIRED MOABS WILL BEGIN BY PREPARATION OF DERIVATIVES OF THE STEROIDS AT SELECTED SITES CHOSEN TO MAXIMIZE CHANCES THAT RESULTING ANTIBODIES WILL BE SPECIFIC.THE DERIVATIVES WILL BE CONJUGATED TO KEYHOLE LIMPET HEMOCYANIN, AND IMMUNIZATION WILL BE PERFORMED IN VIVO USINGBALB/CBYJ MICE. FUSION, SCREENING, SELECTION, AND ANALYSIS WILL INVOLVE PROCEDURES TO ENHANCE THE PROBABILITY OF OBTAINING HIGH-AFFINITY MOABS. PHASE II RESEARCH WILL INVOLVE EXTENDING THE APPROACH TO DEVELOP AND CHARACTERIZE SIMILAR MOABS TO OTHER STEROIDS, PEPTIDE HORMONES AND/OR GROWTH FACTORS, AND THEIR RECEPTORS AS THEY BECOME AVAILABLE. CHOSEN AND SUBCLONED MOABS WILL THEN BE PRODUCED IN QUANITITY IN VITRO USING A CUSTOMIZED IN-HOUSE BIOREACTOR SYSTEM.