THE AIM OF THIS PROJECT IS TO DEMONSTRATE A SENSITIVE HOMOGENEOUS CHEMILUMINESCENCE IMMUNOASSAY FOR METHOTREXATE USING THE PRINCIPLE OF CHEMILUMINESCENCE ENERGY TRANSFER.

THE AIM OF THIS PROJECT IS TO DEMONSTRATE A SENSITIVE HOMOGENEOUS CHEMILUMINESCENCE IMMUNOASSAY FOR METHOTREXATE USING THE PRINCIPLE OF CHEMILUMINESCENCE ENERGY TRANSFER. THIS WILL LEAD TO A TECHNIQUE FOR THE DETECTION OF ANTI- NEOPLASTIC DRUGS REQUIRING HIGHLY SENSITIVE ASSAYS. THE NEWASSAY WILL BE SIMPLER, FASTER, SAFER, AND CHEAPER THAN CONVENTIONAL METHODS. METHOTREXATE WILL BE COVALENTLY COUPLED TO AMINOBUTYLETHYL ISOLUMINOL WITHOUT LOSS OF IMMUNOLOGICAL OR CHEMILUMINESCENCE ACTIVITY. IT WILL THEN BE USED IN CONJUNCTION WITH FLUORESCENT LABELED ANTIBODIES AND THE PRINCIPLE OF ENERGY TRANSFER TO DEVELOP SENSITIVE HOMOGENEOUS (NON-SEPARATION) ASSAYS FOR METHOTREXATE. AFTEROPTIMIZATION, THIS ASSAY WILL BE COMPARED WITH A COMMERCIAL ASSAY. THE RESEARCH IS EXPECTED TO PROVIDE A HOMOGENEOUS METHOTREXATE ASSAY WITH LOWER DETECTION LIMITS, AND STABLE REAGENTS. THE REMOVAL OF THE SEPARATION STEP WILL ALSO LEADTO INCREASED PRECISION AND ACCURACY. THE WORK WILL LEAD TO PHASE II PROTOTYPE INSTRUMENT DESIGN AND PRODUCTION OF ANTIBODIES AND CLINICAL TRIALS FOR ASSAYS OF METHOTREXATE AND OTHER ANTINEOPLASTIC AND THERAPEUTIC DRUGS.