STABILIZATION OF ANTIBODIES FOR ELISA ASSAYS

THIS RESEARCH WILL DEVELOP METHODS TO SIMULTANEOUSLY IMMOBILIZE AND COVALENTLY CROSS-LINK ANTIBODIES OR ANTIGENS ONTO POLYSTYRENE TUBES OR MICROTITER PLATES. THE COVALENTLYIMMOBILIZED ANTIBODIES WILL BE STABILIZED BY MULTIPOINT COUPLING TO HYDROPHILIC POLYMERS ON THE POLYSTYRENE SURFACE. THUS, THE ANTIBODIES WILL BE PROTECTED FROM THE HYDROPHOBIC POLYSTYRENE AND CROSS-LINKED TO ACHIEVE STABILIZATION. BY ACTIVATING LARGE NUMBERS OF FUNCTIONAL GROUPS ON THE COATING POLYMERS, AN AMPLIFICATION OF THE IMMOBILIZATION CAPACITY ON THE POLYSTYRENE SURFACE IS EXPECTED, RESULTING IN GREATER ANTIGEN-BINDING CAPACITIES. OTHER EXPECTED BENEFITS OF THIS POLYSTYRENE COATING TECHNIQUE ARE REDUCED VARIABILITY, REDUCED NONSPECIFIC ADSORPTION, INCREASED SENSITIVITY, AND PERHAPS FASTER BINDING RATES. THIS TECHNIQUE WILL BE DEVELOPED AND ITS FEASIBILITY DEMONSTRATED WITH AN ELISA ASSAY FOR COTININE, A METABOLITE OF NICOTINE. COTININE IS THE MOST RELIABLE DETERMINANT OF USE OF OR EXPOSURE TO TOBACCO.