An improved preparation of antibody-coated polystyrene beads for sandwich enzyme immunoassay.
An improved preparation of antibody-coated polystyrene beads for sandwich enzyme immunoassay of human thyroid-stimulating hormone (TSH) was described. Rabbit anti-TSH IgG was purified by eluting at pH 2.5 from a TSH-Sepharose column, diluted 3 or 9 fold with normal rabbit IgG and used for coating polystyrene beads by physical adsorption. In a sandwich enzyme immunoassay of TSH using rabbit (anti-TSH) Fab'-beta-D-galactosidase conjugate, beta-D-galactosidase activities specifically bound to thus prepared polystyrene beads in the presence of TSH was 2.8-6.3 fold higher than those bound to polystyrene beads coated with anti-TSH IgG before purification. A similar effect was observed when guinea pig anti-pork insulin IgG, rabbit (anti-human IgE) IgG and goat (anti-human IgE) IgG were treated at pH 2.5. This improvement may be based on a conformational change of Fc in IgG molecule which was caused by the treatment at pH 2.5. Other sandwich immunoassays such as fluoro- and radio-immunoassays may also be improved in the same way.
D000818:Animals; D000906:Antibodies; D005696:Galactosidases / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G; D007328:Insulin / Q000276:immunology; D009953:Ornithine-Oxo-Acid Transaminase / Q000276:immunology; D011137:Polystyrenes / Q000494:pharmacology*; D011817:Rabbits; D013972:Thyrotropin / Q000276:immunology
D000906:Antibodies; D007074:Immunoglobulin G; D007328:Insulin; D011137:Polystyrenes; D013972:Thyrotropin; D009953:Ornithine-Oxo-Acid Transaminase; D005696:Galactosidases