Diffusion-in-gel thin layer immunoassay (DIG-TIA): optimal conditions for quantitation of antibodies.
A new technique for the quantitation of antigen-antibody reactions, diffusion-in-gel thin layer immunoassay (DIG-TIA), has been developed. The principle of DIG-TIA is that antibodies are allowed to form a concentration gradient by radial diffusion in an agar gel poured on top of an antigen-coated plastic surface. After removal of the gel and visualisation by means of condensation of water vapour on the plastic surface, the antigen-antibody reactions appear as zones of increased wettability. The concentration of antigen used for coating, pH of the agar, incorporation of Tween 20 in the agar, size of diffusion basins, time of diffusion, and reinforcement by anti-immunoglobulin have been studied with regard to their influence on sensitivity and precision in the detection of antibodies with DIG-TIA. A photographic technique for permanent recording of wettability patterns is also described.
D000704:Analysis of Variance; D000818:Animals; D000906:Antibodies; D000888:Antibodies, Anti-Idiotypic; D000937:Antigen-Antibody Reactions; D000941:Antigens; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000494:pharmacology; D007118:Immunoassay / Q000379:methods; D005779:Immunodiffusion / Q000379:methods; D007158:Immunologic Techniques; D011136:Polysorbates / Q000494:pharmacology; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000276:immunology; D013997:Time Factors
D000906:Antibodies; D000888:Antibodies, Anti-Idiotypic; D000941:Antigens; D007106:Immune Sera; D011136:Polysorbates; D012710:Serum Albumin, Bovine