Quantitative measure of etanercept.
Promonitor®-ETN is an enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of free etanercept (ETN) in human serum samples. The amount of ETN, when considered in conjunction with anti-ETN antibodies (Promonitor®-anti-ETN Ref. 5120230000) and clinical findings, like disease activity, can contribute valuable information to the physician to assess the efficacy of therapies with ETN. It has been described that patients not responding to etanercept obtain lower etanercept concentrations compared with responding patients.
Promonitor®-ETN is a sandwich ELISA to test up to 40 patients. The microwell strips are provided pre-coated with an anti-ETN human F(ab´)2 fragment. Diluted calibrators, controls and diluted patient samples are added to separate wells, allowing ETN present to bind to pre-immobilized anti-ETN antibodies. Unbound sample is washed away, and a second HRP-labeled anti-ETN monoclonal antibody is added to each well. A second incubation step allows the HRP-labeled anti-ETN antibody to bind to the ETN that has become attached to the microwells. After washing away the excess of unbound HRP-labeled anti-ETN antibody, the remaining enzyme activity is measured by adding a chromogenic substrate and measuring the intensity of the color that develops in a spectrophotometer. The signal obtained is proportional to the amount of drug in the patient sample.
Optical densities (OD) of the calibrators are plotted to construct the ETN calibration curve. A four parameter logistic (4PL) model is used to fit the curve. Positive and Negative Controls are used to monitor assay performance. ETN concentration is calculated by interpolating sample OD in the calibration curve. Warnings for each sample are optionally included; this setting can be configured here.
If the entire plate was read then supply the measured value for each well. Alternatively, if only specific parts of the plate were read then list only those measured values. In this case an additional step is required: under the Microplate section below, select or create a layout which defines which wells were read and which were not. More...
A zero concentration value has been specified with a logarithmic x axis.
This is valid but means that you will not see the zero standard on the chart, also in certain cases it may result in a skewed fit. One alternative is to use a suitably low value, such as 0.01 instead of zero. More...
Promonitor® is a trademark of Proteomika, S.L.
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