A project is proposed to develop methods to simultaneously immobilize and covalently crosslink antibodies or antigens onto polystyrene tubes or microtiter plates. The covalently immobilized 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 crosslinked to achieve stabilization. By activating large numbers of functional groups on the coating polymers, an amplification of the immobilization capacity on the polystyrene surface. Thus, the antibodies will be protected from the hydrophobic polystyrene and crosslinked to achieve stabilization. By activating large numbers of functional groups on the coating polymers, an amplification of the antigen binding capacities. Other expected benefits of this proposed 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 for nicotine. Cotinine is the most reliable determinant of use of or exposure to tobacco.
