IMMUNOASSAY FOR HEMOGLOBIN A1C WITH MONOCLONAL ANTIBODY

THIS PROJECT WILL DEVELOP A SIMPLE, SPECIFIC, AND RAPID IMMUNODIAGNOSTIC TEST TO MEASURE THE AMOUNT OF HEMOGLOBIN A1C IN HUMAN ERYTHROCYTES. THE TEST WILL BE AN ELISA CONSTRUCT THAT EMPLOYS A MONOCLONAL ANTIBODY THAT SPECIFICALLY RECOGNIZES HEMOGLOBIN A1C, AN INDEX OF THE AMBIENT BLOOD GLUCOSE CONCENTRATIONS OVER THE PRECEDING 2 TO3 MONTHS. CLINICAL INVESTIGATIONS HAVE ESTABLISHED THAT PERIODIC ASSESSMENT OF THE PREVAILING BLOOD GLUCOSE LEVEL IS ESSENTIAL TO ESTABLISHING AND MAINTAINING CONTROL OF DIABETES. THE MAINSTAY OF MONITORING DIABETIC CONTROL IS MEASUREMENT OF THE BLOOD GLUCOSE CONCENTRATION AND OF THE EXTENT TO WHICH CERTAIN CIRCULATING PROTEINS HAVE BECOME NONENZYMATICALLY GLYCATED. HEMOGLOBIN A1C AND GLYCOHEMOGLOBIN MEASUREMENTS ARE WIDELY USED AS A MEANS TO OBJECTIVELY EVALUATE AMBIENT GLUCOSE CONTROL OVER 2- TO 3-MONTH PERIODS. HOWEVER, CURRENT METHODS FOR THEIR MEASUREMENT ARE TIME CONSUMING, CUMBERSOME, NONSPECIFIC, AND HAVE UNACCEPTABLE COEFFICIENTS OF VARIATION. THESE PROBLEMS COULD BE SURMOUNTED IF A MONOCLONAL ANTIBODY THAT REACTS ONLY WITH HEMOGLOBIN A1C WERE AVAILABLE. THIS PROJECT WILL USE A UNIQUE AND HIGHLY SPECIFIC MONOCLONAL ANTIBODY THAT RECOGNIZES THE N-TERMINAL GLYCATED EPITOPES RESIDING IN HEMOGLOBIN A1C BUT NOT IN OTHER PROTEINS AND THAT DOES NOT REACT WITH (UNGLYCATED) HEMOGLOBIN A DEGREE TO DEVELOP A RELIABLE AND SPECIFIC IMMUNODIAGNOSTIC METHOD TO QUANTITATE HEMOGLOBIN A1C IN HUMAN BLOOD.