TRI-ELISA FOR SIMULTANEOUS ANALYSIS OF THREE ANALYTES

THE ENZYME-LINKED IMMUNOSORBENT ASSARY (ELISA) IS A MAJOR TOOL FOR DETECTING INFECTIOUS AGENTS, PROTEINS, AND DRUGS IN BIOLOGICAL FLUIDS, OR CONTAMINANTS IN ENVIRONMENTAL SAMPLES. THE MAJORITY OF ELISA'S ARE DESIGNED TO DETECT A SINGLE ANALYTE. WE PROPOSE HERE TO DEVELOP A THREE-ENZYME ELISA SYSTEM IN WHICH THREE ANALYTES CAN BE QUANTITATED CONCURRENTLY FROM A SINGLE SAMPLE. WE HAVE IDENTIFIED THREE ENSYMES/SUBSTRATES THAT HAVE MINIMAL OVERLAP AT THREE SEPARATE WAVELENGTHS CORRESPONDING TO THEIR RESPECTIVE MAXIMUM ABSORBANCE. DURING PHASE I, WE WILL DEFINITIVELY IDENTIFY THE THREE ENZYMES AND MULTIPLE SUBSTRATE SOLUTIONS THAT WILL WORK SIMULTANEOUSLY, CONJUGATE EACH ENZYME TO A SEPARATE ANTIBODY, DEVELOP INDIVIDUAL ELISA'S FOR EACH ANALYTE, AND TEST THE THREE ANALYTE SYSTEMS CONCURRENTLY. THE ELISA'S WILL BE DEVELOPED USING A FUNGUS (CANDIDA ALBICANS), A VIRUS (HERPES SIMPLEX VIRUS, TYPE 1), AND A BACTERIA (PSEUDOMONAS AERUGINOSA). THE EXPECTED RESULTS OF THIS PHASE I PROJECT WILL DEMONSTRATE FEASIBILITY AND PROVIDE THE BASIS FOR CONTINUATION INTO A PHASE II PROJECT.