MONOCLONAL ANTIBODES TO EPSTEIN-BARR VIRUS PEPTIDES

EPSTEIN-BARR VIRUS (EBV) IS ONE OF THE MOST COMMON VIRUSES INFECTING HUMANS. FOLLOWING INITIAL VIRULENT INFECTION AND IMMUNE RESPONSE, EBV USUALLY BECOMES DORMANT AND IS RETAINEDFOR THE LIFE OF THE INDIVIDUAL IN A LATENT STATE. EBV MAY BECOME REACTIVATED BY IMMUNE DEFICIENCIES OR OTHER ENVIRONMENTAL OR PHYSIOLOGICAL FACTORS. TWO OF THE MOST STUDIED ANTIGENS OF EBV ARE EBNA-1 AND EA-D BECAUSE THESE POLYPEPTIDES ARE INDICATORS OF LATENT AND VIRULENT STATES OFTHE VIRUS, RESPECTIVELY. THE PHASE I STUDIES WILL CONCENTRATE ON THESE TWO ANTIGENS. MOLECULAR DIAGNOSTIC SYSTEMS, INC. (MDSI) HAS DEVELOPED MONOCLONAL ANTIBODIES TO EBNA-1 AND EA-D, CHARACTERIZED THESE MONOCLONAL ANTIBODIES FOR SPECIFICITY, AND DEMONSTRATED THAT THESE ANTIBODIES RECOGNIZE THE APPROPRIATEEBV ANTIGENS IN CELLS CONTAINING EBV. THE GOAL OF PHASE I IS TO EVALUATE FURTHER THESE MONOCLONAL ANTIBODIES AS RESEARCH REAGENTS. THE SPECIFIC AIMS ARE TO: (1) DEFINE THE EPITOPES RECOGNIZED BY THE MONOCLONAL ANTIBODIES TO EA-D AND EBNA-1, (2) DEVELOP PURIFICATION METHODS FOR EA-D AND EBNA-1 USING MONOCLONAL ANTIBODIES, AND (3) EVALUATE MONOCLONAL ANTIBODY PURIFICATION METHODS FOR THE ISOLATION OF AUTHENTIC EA-D AND EBNA-1 FROM TISSUE CULTURE CELLS. IN PHASE II, THE SUCCESSFUL METHODS DEVELOPED IN PHASE I WILL BE APPLIED TO OTHER IMPORTANT EBV PROTEINS.