Detection of anti-ovalbumin IgG in serum by immune complex transfer enzyme immunoassay.
An immune complex transfer enzyme immunoassay for anti-ovalbumin IgG in serum is described. Serum-specific antibody was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-ovalbumin conjugate and ovalbumin-peroxidase conjugate. The complex formed by the three components was trapped onto polystyrene balls coated with anti-2,4-dinitrophenyl group IgG, eluted with epsilonN-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with anti-human IgG-gamma-chain. Bound peroxidase activity was determined by fluorometry. This enzyme immunoassay was 300- to 1,000-fold more sensitive and more reliable than the enzyme-linked immunosorbent assay (ELISA). Anti-ovalbumin IgG was detected in 100% of healthy subjects using this method while only 14% were detected by ELISA.
Hirota
K
K
Division of Maternal and Child Health Science, National Institute of Health and Nutrition, Tokyo, Japan. khirota@nih.go.jp
Kamashima
T
T
Totani
M
M
D019297:2,4-Dinitrophenol / Q000276:immunology; D000818:Animals; D000936:Antigen-Antibody Complex; D000941:Antigens / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood*; D007142:Immunoglobulin gamma-Chains; D008863:Microspheres; D010047:Ovalbumin / Q000276:immunology*; D009195:Peroxidase / Q000276:immunology; D011137:Polystyrenes; D011817:Rabbits; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000276:immunology
D000936:Antigen-Antibody Complex; D000941:Antigens; D007074:Immunoglobulin G; D007142:Immunoglobulin gamma-Chains; D011137:Polystyrenes; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D009195:Peroxidase; D019297:2,4-Dinitrophenol