Pyrophosphatase-based enzyme-linked immunosorbent assay of total IgE in serum.
Human IgE was purified to near homogeneity by a two-step procedure consisting of immunoaffinity chromatography and high-performance liquid chromatography. Mouse hybridoma cell lines secreting antibodies against IgE were generated. Monoclonal and polyclonal antibodies were used to develop a "sandwich"-type ELISA for determining total IgE in human serum. Inorganic pyrophosphatase (EC 3.6.1.1), an enzyme having a high turnover number, was used as the label. The mean analytical recovery of our ELISA was 95.2% and the results showed good linear correlation with an established RIA of IgE. We found pyrophosphatase to be a good alternative label for use in immunoassays.
Clinical Research Laboratory, Turku, Finland.
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Peuravuori
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Korpela
D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010744:Phosphoric Monoester Hydrolases; D011786:Quality Control; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D007073:Immunoglobulin E; D010744:Phosphoric Monoester Hydrolases