AN IMMUNOASSAY FOR GLYCOSYLATED HEMOGLOBIN

DIAGNOSIS AND MANAGEMENT OF DIABETES MELITUS IS FACILITATED MEASUREMENTS OF GLYCOSYLATED HEMOGLOBIN IN PATIENT SERUM. RELATIVE LEVELS OF THIS MOLECULE ARE ELEVATED IN THE DIABETIC PATIENT. THE PRESENTLY AVAILABLE METHODS FOR THESE MEASUREMENTS ARE TIME CONSUMING AND REQUIRE CONSIDERABLE TECHNICAL EXPERTISE. IN THE WORK PROPOSED HERE, WE PLAN TO GENERATE MONOCLONAL ANTIBODIES SPECIFIC FOR THE GLYCOSYLATED RESIDUE ON GLYCOSYLATED HEMOGLOBIN. THIS WILL BE ACCOMPLISHED BY IMMUNIZING MICE THREE DIFFERENTTYPES OF IMMUNOGEN: PURIFIED GLYCOSYLATED HEMOGLOBIN, PEPTIDES DERIVED FROM GLYCOSYLATED HEMOGLOBIN AND THEN CONJUGATED TO IMMUNOGENIC CARRIER PROTEINS, AND PURIFIED GLYCOPEPTIDES CONJUGATED TO LIPIDS. THESE WILL BE INJECTED IN CONCERT WITH A VARIETY OF ADJUVANTS TO GENERATE MURINE HYBRIDOMAS SECRETING MONOCLONAL ANTIBODIES CAPABLE OF DISTINGUISHING THE TWO FORMS OF HEMOGLOBIN. THESE ANTIBODIES WILL BE USED TO DEVELOP A "SANDWICH" ELISA FOR MEASURING GLYCOSYLATED HEMOGLOBIN IN PATIENT SAMPLES. IN PHASE II STUDIES, THE DEVELOPMENT OF THIS ASSAY WILL BE COMPLETED, OPTOMOIZED AND VALIDATED, AND EVALUATUATED USING CLINICAL MATERIAL.