A novel enzyme immunoassay of anti-insulin IgG in guinea pig serum.
A novel enzyme immunoassay of anti-insulin IgG in guinea pig serum is described. Guinea pig anti-insulin serum diluted with nonspecific guinea pig serum was incubated with dinitrophenyl biotinyl nonspecific rabbit IgG-insulin conjugate and a rabbit (anti-dinitrophenyl bovine serum albumin) IgG-coated polystyrene ball. After washing to eliminate nonspecific guinea pig IgG in the diluted serum, the polystyrene ball was incubated with dinitrophenyl-L-lysine to elute the complex of anti-insulin IgG and the conjugate. The eluate was incubated with an avidin-coated polystyrene ball. Finally, the amount of guinea pig anti-insulin IgG in the complex trapped onto the avidin-coated polystyrene ball was measured by incubation with rabbit (anti-guinea pig IgG) Fab'-peroxidase conjugate. This enzyme immunoassay was 10,000-fold more sensitive than the conventional enzyme immunoassay using insulin-coated polystyrene ball and rabbit (anti-guinea pig IgG) Fab'-peroxidase conjugate.
D000818:Animals; D001710:Biotin / Q000276:immunology; D004140:Dinitrophenols / Q000276:immunology; D006168:Guinea Pigs; D006241:Haptens / Q000276:immunology; D007106:Immune Sera / Q000032:analysis; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis*; D007328:Insulin / Q000276:immunology*; D008239:Lysine / Q000031:analogs & derivatives / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000276:immunology
D004140:Dinitrophenols; D006241:Haptens; D007106:Immune Sera; D007074:Immunoglobulin G; D007328:Insulin; C034638:dinitrophenyl-bovine serum albumin; C007493:epsilon-dinitrophenyllysine; D012710:Serum Albumin, Bovine; D001710:Biotin; D008239:Lysine