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Quantitation of serum IgE by using chimeras of human IgE receptor and avian immunoglobulin domains

Braren, Ingke, Greunke, Kerstin, Pilette, Charles, Mempel, Martin, Grunwald, Thomas, Bredehorst, Reinhard, Ring, Johannes, Spillner, Edzard, Ollert, Markus
Analytical biochemistry 2011 v.412 no.2 pp. 134-140
antibodies, asthma, birds, blood serum, chimerism, enzyme-linked immunosorbent assay, humans, immunoblotting, immunoglobulin E, immunoglobulin Y, monitoring, patients, therapeutics
Anti-IgE therapeutics represent an efficient approach in the management of IgE-mediated allergic asthma. However, monitoring the reduction of IgE levels into a therapeutically efficient range requires the determination of residual serum IgE. We established an analytical approach to distinguish free and anti-IgE complexed serum IgE based on soluble derivatives of the human high-affinity IgE receptor. Soluble receptor derivatives represent an ideal means to analyze receptor antagonism by any ligand or blocking antibody. Therefore, the FcεRI ectodomain was fused with avian IgY constant domains that circumvent susceptibility to interference phenomena and improve assay performance. After production in HEK293 cells, subsequent characterization by enzyme-linked immunosorbent assay and immunoblotting confirmed the suitability of avian IgY constant domains for immobilization and detection purposes. To provide further insights into the different IgE reactivities, free allergen-specific IgE was also determined. Monitoring of sera from omalizumab-treated patients during the course of therapy revealed the applicability for assessment of omalizumab-complexed versus noncomplexed serum IgE. These parameters may allow correlation to clinical responses during anti-IgE therapy with the perspective of biomonitoring.