Detection of Mouse IgE by CELISA

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Abstract
The detection and quantitation of mouse IgE is usually impaired by the difficulty to obtain reliable antibody reagents which are fully specific for the ε chain – and reactive enough – to be used in an enzyme-linked immunosorbent assay (ELISA). An ELISA on cells (CELISA) was developed for the detection of mouse IgE, using rat basophilic leukemia (RBL) cells. It is based on the high affinity of the receptors for the Fc of IgE (FcεR) displayed on the surface of the RBL cells. Since the ε chain specific recognition is achieved by the biological receptor of IgE, the detection of cell-bound IgE does not need the use of ε chain specific antibodies. Instead, one can use any enzyme-coupled antibody capable to recognize the IgE through its light-chain epitopes. Interestingly, when the IgE bound to the RBL cells has a known specificity, it can be detected through its paratopes using the cognate antigen coupled to an enzyme.