Clinical Chemistry
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Clinical Chemistry 39: 846-851, 1993;
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Clinical Chemistry, Vol 39, 846-851, Copyright © 1993 by American Association for Clinical Chemistry

Pyrophosphatase-based enzyme-linked immunosorbent assay of total IgE in serum

H Peuravuori and T Korpela
Clinical Research Laboratory, Turku, Finland.

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.




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Copyright © 1993 by the American Association for Clinical Chemistry.