Golimumab (Simponi ®) ELISA Assay Kit


The Eagle Biosciences Golimumab (Simponi ®) ELISA Assay Kit is an enzyme immunoassay for the analytical determination of free Golimumab in serum and plasma samples. This assay utilizes the target molecule (TNF-α) as the binding reagent on the microtiter plate so this assay will measure any other therapeutic that targets TNF-α including Adalimumab, Etanercept, and Infliximab.  The Golimumab ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Golimumab (Simponi®)ELISA Assay Kit

For Research Use Only

Size: 1 x 96 wells
Sensitivity: 5 ng/mL
Dynamic Range: 6-200 ng/mL
Incubation Time: 2 hours
Sample Type:  Serum, Plasma
Sample Size: 10 µL

Additional Information

Assay Background

Golimumab (Simponi ®) is a human monoclonal antibody that binds to both the soluble and transmembrane bioactive forms of human TNFα. This interaction prevents the binding of TNFα to its receptors, thereby inhibiting the biological activity of TNF. Golimumab has been proven effective in the treatment of Rheumatoid Arthritis (RA), Ankylosing Spondylitis (AS), Psoriatic Arthritis (PsA) or Ulcerative Colitis (UC). Antibodies to Golimumab were detected in 57 (4%) of Golimumab -treated patients across the Phase 3 RA, PsA, and AS trials through Week 24. Similar rates were observed in each of the three indications. Patients who received Golimumab with concomitant Methotrexate (MTX) had a lower proportion of antibodies to Golimumab than patients who received Golimumab without MTX (approximately 2% versus 7%, respectively). Of the patients with a positive antibody response to Golimumab in the Phase 2 and 3 trials, most were determined to have neutralizing antibodies to Golimumab as measured by a cell-based functional assay. The data from the literature demonstrated that Anti-Drug Antibody positivity was significantly associated with low Golimumab levels and poor therapeutic response. The positive correlation between serum drug trough levels and therapeutic response indicates that drug monitoring could be useful for optimising the dosing of biologics in a personalised therapy strategy.

Identification of biomarkers for (non-)response and risk factors for adverse drug reactions that might be related to serum concentrations and maintaining the effective concentration of Golimumab in order to potentially avoid some side effects with a reliable method might be beneficial.

Assay Principle

This Eagle Biosciences Golimumab ELISA Assay Kit is based on sandwich type ELISA. Diluted standards and samples (serum or plasma) are incubated in the microtiter plate coated with recombinant human TNF-α (rh TNF-α). After incubation, the wells are washed. A horseradish peroxidase (HRP) conjugated anti-human IgG monoclonal antibody is added and binds to the Fc part of Golimumab pre-captured by the rhTNF-α on the surface of the wells. Following incubation, the wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The color developed is proportional to the amount of free Golimumab in the sample or standard. Results of samples can be determined by using the standard curve.

Assay Procedure

  1. Pipette 100 µL of Assay Buffer into each of the wells to be used.
  2. Pipette 75 µL of each 1:10 Diluted Standard, and 1:50 Diluted Samples into the respective wells of the microtiter plate.
  3. Cover the plate with adhesive seal. Shake plate carefully. Incubate 60 min at room temperature (RT, 20-25°C).
  4. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  5. Pipette 100 μL of Enzyme Conjugate (HRP-anti human IgG mAb) into each well.
  6. Cover plate with adhesive seal. Shake plate carefully. Incubate 30 min at RT.
  7. Remove adhesive seal. Aspirate or decant the incubation solution. Wash the plate 3 X 300 μL of Diluted Wash Buffer per well. Remove excess solution by tapping the inverted plate on a paper towel.
  8. Pipette 100 µL of Ready-to-Use TMB Substrate Solution into each well.
  9. Incubate 15 min at RT. Avoid exposure to direct sunlight.
  10. Stop the substrate reaction by adding 100 µL of Stop Solution into each well. Briefly mix contents by gently shaking the plate. Color changes from blue to yellow.
  11. Measure optical density (OD) with a photometer at 450 nm (Reference at OD620 nm is optional) within 15 min after pipetting the Stop Solution.

Typical Standard Curve


There is no cross reaction with any other proteins present in naïve serum. Because the solid phase is coated with rhTNF-α, other therapeutic anti-TNF antibodies cause full cross reaction. However, a quantification of other therapeutic antibodies is possible only by using the drug-specific standards, which are available as separate kits from Eagle Biosciences.


Product Manual



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