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Isolation of IgE from Reaginic Rat Serum

From the Section on Chemical Immunology, Arthritis and Rheumatism Branch, National Institute of Arthritis, Metabolism and Digestive Diseases, National Institutes of Health, Bethesda, Maryland 20014

Abstract

IgE from the serum of rats immunized with Nippostrongylus brasiliensis was substantially purified. As measured by passive cutaneous anaphylaxis (PCA) activity an overall yield of 45% was reproducibly achieved by a combination of ammonium sulfate precipitation, gel and ion-exchange chromatography, and immunoadsorbents. The iodinated final product was 80% precipitable by specific antisera to IgE, and in radio-immunoelectrophoresis with antiserum to light chains gave only a single line corresponding to the line given by antisera to IgE. No lines could be detected with an antiserum to normal serum proteins devoid of antibodies to light chains. About 80% of the IgE related counts were shown to bind specifically to neoplastic rat basophils. On the basis of these results it was calculated that 18 ng of rat IgE are adequate to give a detectable PCA response.

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