RecoveryELISA IgE / Omalizumab Kit (RIO), 96 Well

The recoveryELISA IgE / Omalizumab (RIO) is a test kit (Research Use Only) for the simultaneous quantitative determination of free IgE, IgE neutralization rate and the available therapeutic antibody Omalizumab in human serum samples during therapy. It is a manual, non-automated kit for the determination of 7 samples.
With the recoveryELISA IgE / Omalizumab Kit (RIO) the following parameters will be obtained:
1. free IgE concentration
2. IgE neutralization rate
3. level of available Omalizumab in the sera of the patients, during therapy.
The test result does not represent a recommendation for dose adjustments or treatment. Evaluation of the test result is the responsibility of the supervising doctor.
The recoveryELISA is an immunological quantitative detection method based on a sandwich ELISA. In contrast to a classic sandwich ELISA, a two-dimensional calibration is carried out.
The following calibrations are performed:
IgE levels without and with additional Omalizumab against extinction (optical density)
Omalizumab levels against IgE recovery
The recoveryELISA is performed in a 96-well microplate. The wells of the microplate are pre-coated with a capture antibody against human IgE. A simultaneous incubation of the IgE (with and without the addition of Omalizumab), the samples and the detection conjugate (peroxidase conjugate of Omalizumab) is carried out. The incubation time is 16 to 22 h at 2 to 8°C. After washing, the colour substrate TMB (Tetramethylbenzidine) is added.
The enzymatic colour reaction is stopped using sulphuric acid solution and the colour change from blue to yellow is optically detected using a wavelength of 450 nm (reference value 620 nm). The optical density (OD) of the reaction product is measured with a suitable microplate reader.
With the aid of the two calibrations, an evaluation procedure can be carried out to determine the concentrations of free IgE and Omalizumab in the patient samples using the measured OD values. Determination of the therapeutic antibody is based on the principle that the presence of Omalizumab in patient samples leads to a systematic reduction in the recovery of IgE.
The evaluation is performed using non-linear regression (Marquardt-Levenberg algorithm), the Michaelis-Menten model from enzyme kinetics and the Langmuir isotherm from surface binding.

Measurement Range for undiluted samples:
IgE Levels in samples without Omalizumab 1 – 2000 IU / mL
IgE Levels in samples with Omalizumab 1 – 500 IU / mL
Omalizumab level: 0.4 – 80 µg / mL

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