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Detection of True IgE-expressing Mouse B Lineage Cells
Published on: December 1, 2014
Estimation of IgE antibody with a nonisotopic technique with no interference of isotypic antibodies
T M Lee1, L C Grammer, M A Shaughnessy
1Department of Medicine, Northwestern University Medical School, Chicago, IL 60611.
A modified ELISA was developed to estimate antigen-specific IgE antibody. IgE against ragweed antigen E (AgE) was the model system, and paired pre- and postimmunotherapy sera were evaluated because the latter had been demonstrated to contain significant amounts of IgG antibody. Patients' serum IgE was bound to the surface of wells of a polystyrene microtiter plate previously coated with monoclonal murine antihuman IgE. After washing, AgE conjugated with alkaline phosphatase was added and was specifically bound by IgE. Optical density was recorded after addition of enzyme substrate to the wells and was proportional to the concentration of IgE directed against ragweed AgE in the serum samples. In contrast to many other assay techniques for antigen-specific IgE, interference by specific antibody of other classes cannot occur with this method. Results correlated well with those of a previously described solid-phase radioimmunoassay that required myeloma IgE.
A modified ELISA was developed to estimate antigen-specific IgE antibody. IgE against ragweed antigen E (AgE) was the model system, and paired pre- and postimmunotherapy sera were evaluated because the latter had been demonstrated to contain significant amounts of IgG antibody. Patients' serum IgE was bound to the surface of wells of a polystyrene microtiter plate previously coated with monoclonal murine antihuman IgE. After washing, AgE conjugated with alkaline phosphatase was added and was specifically bound by IgE. Optical density was recorded after addition of enzyme substrate to the wells and was proportional to the concentration of IgE directed against ragweed AgE in the serum samples. In contrast to many other assay techniques for antigen-specific IgE, interference by specific antibody of other classes cannot occur with this method. Results correlated well with those of a previously described solid-phase radioimmunoassay that required myeloma IgE.
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