TNF-alpha Neutralization Rate Adalimumab ELISA Assay

$1,095.00

The TNF-alpha Neutralization Rate Adalimumab ELISA Assay is an in-vitro diagnostic agent for the simultaneous quantitative determination of free TNFα, the TNFα neutralization rate and the available therapeutic antibody Adalimumab in human serum samples. It consists of a manual, non-automated kit for the determination of 7 samples. The recoveryELISA RTA is for research use only and not to be used in diagnostic procedures.

TNF-alpha Neutralization Rate Adalimumab ELISA Assay

The TNF-alpha Neutralization Rate Adalimumab ELISA Assay is For Research Use Only

Size: 1x96wells
Sensitivity: Adalimumab: 0.7 µg/mL; TNF alpha: 4ng/ml
Dynamic Range: Adalimumab: 0.7 – 30 µg/mL; TNF alpha in sample without Adalimumab: 4-400ng/ml;TNF alpha in sample with Adalimumab: 4-160ng/ml
Incubation Time: Overnight
Sample Type: Serum
Sample Size: 60 µL

Controls Included


Assay Background

Tumour necrosis factor (TNFα) is a signalling substance of the immune system which can cause, among other things, the formation of inflammatory mediators in cells. In inflammatory diseases such as rheumatoid arthritis, there is overproduction of TNFα, which can be inhibited by using therapeutic antibodies.

Adalimumab is an IgG1 human monoclonal antibody which is used as a TNFα inhibitor. Adalimumab binds specifically to the TNFα protein and neutralises the biological effect of TNFα.

Adalimumab is approved for the treatment of the following diseases: rheumatoid arthritis, psoriatic arthritis, polyarticular juvenile idiopathic arthritis, ankylosing spondylitis, Crohn’s disease and ulcerative colitis, but is also used “off-label” for other conditions such as sarcoidosis. In therapy using adalimumab, contraindications such as hypersensitivity to adalimumab and also serious infections, sepsis, tuberculosis and cases of opportunistic infections have been noted. Weakening of the body’s defence is produced by the immunosuppressive effect of adalimumab. Excessive neutralisation of TNFα during treatment with adalimumab increases the risk of developing opportunistic infections. In view of this, monitoring the treatment antibody levels and degree of neutralization of TNFα would appear useful during therapy using the TNFα inhibitor adalimumab. In comparison with the tests available on the market, the recoveryELISA TNFα Neutralization Rate/Adalimumab (RTA) Kit can be used to simultaneously determine the free TNFα target antigen, the available therapeutic antibody adalimumab and its ability to neutralise TNFα.


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Additional Information

Assay Principle


RecoveryELISA is performed in a 96-well microplate. The wells of the microplate are pre-coated with a specific capture antibody against human TNFα that binds free TNFα from the patient sample. There occurs a simultaneous incubation of the TNFα levels (with and without the addition of Adalimumab) of the samples and of the detection conjugate (peroxidase conjugate of an anti-TNFα antibody). Incubation takes place over 16 to 22 hours at 2 to 8°C. After washing, the colour substrate TMB (tetramethylbenzidine) is added to the wells. After incubation the enzymatic colour reaction is stopped using a sulphuric acid solution. A colour change from blue to yellow occurs that 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 to be performed, an evaluation procedure can be carried out to determine the concentrations of free TNFα and Adalimumab in the patient samples using the measured OD values. Determination of the therapeutic antibody is based on the principle that the presence of Adalimumab in patient samples leads to a systematic reduction in the recovery of TNFα.

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References


  1. Strohner P: Patent: EP 1957980B1 Immunoassay for the simultaneous immunochemical determination of an analyte (antigen) and a treatment antibody targeting the analyte in samples (Recovery immunoassay)
  2. Strohner P, Sarrach D, Reich JG, Becher G, Staatz A, Schäfer A, Steiß J-O, Häupl T: The Recovery-ELISA – a novel Assay Technique for the Therapy Control of Therapeutic Antibodies Am J Respir Crit Care Med 2010;181:A5674
  3. Strohner P, Staatz A, Schäfer A, Becher G, Sarrach D, Reich JG, Steiß J-O, Häupl T: Monitoring of Monoclonal Antibody Therapy with a new Recovery ELISA Assay Technique (RELISA®) ERS Barcelona 2010, Abstract-No: 3399
  4. Strohner P, Staatz A, Sarrach D, Steiß J-O, Becher G: The recoveryELISA – a newly devloped immunoassay for measurement of therapeutic antibodies and the target antigen during antibody therapy Clin Chem Lab Med 2012;50(7):1263-1269

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