A novel immunoassay system and bioseparation process based on thermal phase separating polymers.
Poly-N-isopropylacrylamide (polyNIPAAm), a water-soluble, thermally precipitating synthetic polymer, has been conjugated together with a monoclonal antibody (MAb) and utilized in a novel separation method for an immunoassay. The PolyNIPAAm precipitates out of water above a critical temperature of 31 degres C, enabling a polymer-bound immune complex to be separated from the solution. The principal advantages of this method are that it utilizes a homogeneous incubation for the antigen-antibody reaction, plus, it has the ability to assay large-molecular-weight antigens with sensitivities equivalent to other nonisotopic heterogeneous immunoassays. In addition, since the polymer-immune complex may be reversibly redissolved by cooling, the method may be used both to concentrate the signal and isolate the analyte. This general technique may also be used for a wide variety of separation processes in addition to immunoassays, in which a specific component in a biological fluid, industrial process stream, or body of water is to be isolated for analysis, recovery, or disposal. Thus, product recovery and/or toxin or pollutant removal processes are possible with this methodology.
D000180:Acrylic Resins; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis*; D007145:Immunoglobulin kappa-Chains; D051379:Mice; D013861:Thiocyanates
D000180:Acrylic Resins; D000911:Antibodies, Monoclonal; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D013861:Thiocyanates; C052970:poly-N-isopropylacrylamide; D016650:Fluorescein-5-isothiocyanate