Immunological methods for quantifying free and total serum IgE levels in allergy patients receiving omalizumab (Xolair) therapy.

Omalizumab (humanized-IgG1 anti-human IgE Fc, Xolair) complexes circulating IgE, blocking IgE binding to high affinity epsilon Fc receptors (FcepsilonR1) on mast cells and basophils. Free (non-Omalizumab bound) IgE levels in serum are a measure of effective Omalizumab dosing. The goal of this study was to quantify free (non-Omalizumab-complexed) and total serum IgE levels in asthma patients on Xolair. The concentration of (non-Omalizumab bound) free IgE in human serum was measured using a solid phase immunoenzymetric assay (IEMA) in which IgE was captured from serum with monoclonal anti-human IgE (clone HP6061) and detected with labeled-FcepsilonR1alpha. In a companion total human serum IEMA, IgE was captured from serum with the same anti-human IgE (clone HP6061) and all bound IgE was detected with labeled monoclonal anti-human IgE Fc (clone HP6029). Free and total IgE levels were quantified in pre- and 1 and 3 months post Omalizumab therapy sera from 12 allergic asthma patients. In the absence of Omalizumab, working ranges of the free and total IgE IEMAs were comparable (10-1000 kIU/l), with excellent precision, reproducibility and parallelism. Pre-Omalizumab total and free IgE levels by IEMA were highly correlated (r2=0.99, Y=0.9X+0.32, p<0.001), as were total serum IgE levels by IEMA and ImmunoCAP-250 (r2=0.98, Y=1.1X-0.05, p<0.001, n=33). In vitro reduction of free IgE (>90%) occurred at [Omalizumab:IgE] molar ratios of 2-20. Total IgE levels in 12 asthmatics increased from pre-therapy levels (52-658 kIU/l) by 1.5-5.5-fold at 1 month and 1.7-8.6 fold at 3 months of uninterrupted Omalizumab treatment. Free IgE levels fell by 49%-97% at 1 month and 45%-98% by 3 months of Omalizumab treatment. Free and total IgE levels by IEMA aid in monitoring patients receiving Omalizumab therapy.