FERRITIN-LINKED IMMUNOASSAY: A NONRADIOACTIVE METHOD

THE OBJECTIVE OF THIS PROPOSAL IS TO DEVELOP A NEW NONRADIOACTIVE IMMUNOCHEMICAL ASSAY ^ FERRITIN-LINKED IMMUNOASSAY (FLIA) ^ WHICH MAY REPLACE, AT LEAST PARTIALLY, THE MARKET CURRENTLY MONOPOLIZED BY RIA (RADIOIMMUNOASSAY). FLIA IS COMPARABLE TO RIA IN SIMPLICITY AND IN SENSITIVITY. THE ADVANTAGES OF FLIA INCLUDE LOWER COST, REDUCED HEALTH RISK, AND THE ELIMINATION OF RADIOACTIVE WASTE DISPOSAL. HOWEVER, THE UNIQUE AND INNOVATIVE FEATURE OF FLIA IS THAT THE ANTIGENS ARE LABELED WITH LARGE NUMBERS OF IRON ATOMS WHICH IN TURN ARE STORED IN FERRITIN MOLECULES. USING SIMPLE IRON DETECTION METHODS, THE AMOUNT OF IRON IN THE IMMUNE COMPLEX CAN BE EASILY DETERMINED TO ESTIMATE THE QUANTITY OF AN ANTIGEN AT THE PICOMOLE LEVEL. DURING PHASE I, A PILOT FLIA SYSTEM USING TRINITROPHENYL GROUP (TNP) AS THE ANTIGEN WILL BE PRODUCED. THE SPECIFIC AIMS ARE: 2.TO MAXIMIZE THE IMMUNOPRECIPITATION BETWEEN ANTI-TNP-ANTISERUM AND FERRITIN-LINKED TNP, AND FERRITIN HAS BEEN SHOWN TO FORM CONJUGATES WITH MANY DIFFERENT MOLECULES INCLUDING HORMONES, DRUGS, ETC. THUS, THE ULTIMATE GOAL OF THIS PROPOSAL DURING PHASES II AND III IS TO DEVELOP A VARIETY OF FLIA SYSTEMS FOR COMPOUNDS WHICH ARE OF BIOMEDICAL SIGNIFICANCE.