Dot-immunobinding assay with monoclonal anti-IgE antibodies for the detection and quantitation of human IgE.
This paper describes a dot immunobinding assay for determining total human IgE with a tandem of monoclonal anti-IgE antibodies. Minute quantities of monoclonal anti-IgE antibodies were adsorbed on nitrocellulose discs. IgE bound to this solid phase monoclonal anti-IgE antibody was detected by a second monoclonal antibody conjugated with horseradish peroxidase. Using 4-chloro-1-naphthol as a chromogen results in a stable colour reaction that can be semiquantitatively analysed by the naked eye. The colour intensities of the reaction were also analysed by densitometry, yielding a very reproducible quantitation of human serum IgE. Using a serum dilution of 1:50, IgE could be detected in the range of 12.5-2500 U/ml. Using non-diluted serum samples IgE levels between 0.05-50 U/ml were reproducibly measured. Total serum IgE as determined by this dot assay correlated very well with IgE determinations performed by the commercial PRIST assay.
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006735:Horseradish Peroxidase; D006801:Humans; D007124:Immunoenzyme Techniques; D007073:Immunoglobulin E / Q000032:analysis*; D007202:Indicators and Reagents
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007202:Indicators and Reagents; D007073:Immunoglobulin E; D006735:Horseradish Peroxidase