CONTROL AND STANDARDIZATION OF APOLIPOPROTEIN ASSAYS

IN 1984 AN NIH CONSENSUS CONFERENCE RECOMMENDED COMMUNITY BASED CHOLESTEROL SCREENING PROGRAMS TO EVALUATE HYPERCHOLESTEROLEMIA IN THE CONTEXT OF ITS ASSOCIATION WITH INCREASED HEART ATTACKS. THERE IS A DEVELOPING LITERATURE THAT THE APOLIPOPROTEINS A-1 AND B ARE BETTER MEASURES OF RISK THAN HDL AND LDL CHOLESTEROL WHICH CONTAIN THESE PROTEINS. DEVELOPMENT OF APOLIPOPROTEIN ASSAYS USING MONOCLONAL ANTIBODIES WILL PROVIDE BETTER DIAGNOSIS OF RISK THAN THE LIPOPROTEIN CHOLESTEROL LEVELS. THE OVERALL GOAL OF THIS PROGRAM IS TO DEVELOP, PRODUCE AND MARKET SUCH AN ASSAY. A MAJOR PROBLEM IN THE DEVELOPMENT OF THESE ASSAYS LIES IN THE LACK OF STABLE CONTROLS AND STANDARDS. THEREFORE, THE OVERALL OBJECTIVE OF THIS PHASE I PROJECT IS TO DEVELOP STABLE STANDARDS AND CONTROLS SUITABLE FOR USE IN APOLIPOPROTEIN ASSAYS USING STABLE SOLID PHASE CORES WHICH SIMULATE THE SIZE OF LIPOPROTEINS AND WHICH CONTAIN ON THEIR SURFACE STABLE FRAGMENTS OF APOLIPOPROTEINS CONTAININGTHE EPITOPES OR DETERMINANTS DEFINED BY SPECIFIC MONOCLONAL ANTIBODIES. THE SPECIFIC AIMS INCLUDE: (1) ISOLATING FRAGMENTS OF HUMAN APO A-1 AND APO B; (2) BINDING THESE FRAGMENTS TO 50 NM PARTICLES; AND (3) DEFINING THE STABILITYOF THESE SURROGATE LIPOPROTEINS. PHASE II FUNDING WILL BE REQUESTED TO PREPARE IMMUNOASSAY KITS, CONDUCT CLINICAL TRIALS, AND FOR PRODUCTION SCALEUP. AN IMMUNOASSAY WITH STABLE CONTROLS AND STANDARDS SHOULD IMPROVE DIAGNOSIS OF RISK FOR HEART DISEASE THEREBY REDUCINGMEDICAL COSTS BY EARLY INTERVENTION. THE GENERIC CONTROLS AND STANDARDS DEVELOPED ON THIS PROJECT WOULD BE USEFUL FOR OTHER ASSAYS BENEFITTING THE ENTIRE MEDICAL COMMUNITY AS WELL AS THE PATIENT.