A modified crossed radioimmunoelectrophoretic method for determination of allergen-specific IgG antibodies. Elimination of non-specific binding by use of F(ab')2-fragments of rabbit antibodies and 125I-labelled protein A.
A crossed radioimmunoelectrophoretic (CRIE) method for detection of specific IgG antibodies in patients' sera against horse hair and dander was developed. The unacceptably high non-specific binding encountered when substituting 125I-labelled antihuman IgG for 125I-labelled antihuman IgE in an ordinary CRIE was eliminated by the combined use of 125I-labelled Protein A as detector, and F(ab')2-fragments of the allergen-specific rabbit antibodies. The low background binding thus obtained makes the method useful for detection of specific IgG in sera where the ratio between specific and non-specific IgG is low. Therefore the method should also be applicable to other antigen/allergen systems.
D000818:Animals; D000918:Antibody Specificity; D000939:Epitopes; D006197:Hair / Q000276:immunology; D006736:Horses; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007123:Immunoelectrophoresis, Two-Dimensional; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology*; D007457:Iodine Radioisotopes; D013205:Staphylococcal Protein A / Q000378:metabolism
D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D013205:Staphylococcal Protein A