Certolizumab Pegol mAb-based ELISA


The Certolizumab Pegol mAb-based ELISA is an enzyme immunoassay for the specific and precise determination of free Certolizumab pegol in serum and plasma. The Certolizumab pegol ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Certolizumab Pegol mAb-based ELISA

The Certolizumab Pegol mAb-based ELISA is For Research Use Only

Size: 1 x 96 wells
Sensitivity: 2 ng/mL
Dynamic Range: 6 – 200 ng/mL
Incubation Time: 3 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Name: Cimzia

Assay Principle

This Certolizumab Pegol mAb-based ELISA is based on Certolizumab pegol-specific mouse monoclonal antibody (catcher Ab, ImmunoGuide clone CY). Standards and diluted samples are incubated in the microtiter plate coated with IG-CY mAb. After incubation, the wells are washed. A biotinylated recombinant human tumor necrosis factor alpha (rhTNF-α) is added and binds to the Fab part of Certolizumab pegol. Following incubation, wells are washed and the horseradish peroxidase (HRP)-conjugated streptavidin is added and binds to the biotinylated rhTNF-α. Following incubation, wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The colour developed is proportional to the amount of Certolizumab pegol in the sample or standard. Results of samples can be determined by using the standard curve. Preincubation of Certolizumab pegol with rhTNF-α inhibited the reaction in a concentration dependent manner. Therefore, the ImmunoGuide Certolizumab pegol ELISA (mAb-based) measures the free form of Certolizumab pegol.Pipette 100 µL of Assay Buffer into each of the wells to be used.
Pipette 50 µL of each 1:500 Diluted Standard, and 1:500 Diluted Samples into the respective wells of the microtiter plate.
Cover the plate with adhesive seal. Shake plate carefully. Incubate 60 min at room temperature (RT, 20-25°C).
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.

Related Products

Antibody to Certolizumab Pegol ELISA Kit
Nivolumab (Opdivo) ELISA (mAb-based) Assay Kit
Ustekinumab mAb-based ELISA

Additional Information

Assay Background

Certolizumab pegol (trade name Cimzia®) is a tumor necrosis factor alpha (TNFα) blocker and binds to human TNFα with a KD of 90pM. Certolizumab pegol is a recombinant, humanized antibody Fab’ fragment, with specificity for TNFα, conjugated to an approximately 40kDa polyethylene glycol (PEG2MAL40K). The Fab’ fragment is manufactured in E. coli and is subsequently subjected to purification and conjugation to PEG2MAL40K, to generate Certolizumab pegol. Steady-state concentrations range from 0.5 to 90 µg/mL for a fixed dose of 400 mg of certolizumab pegol.  The ImmunoGuide Certolizumab pegol ELISA  (mAb-based) kit can be efficiently used for measuring free Certolizumab pegol levels in serum and plasma.

Assay Procedure

  1. Pipette 100 μL of Biotinylated TNF into each well.
  2. Cover the plate with adhesive seal. Shake plate carefully. Incubate 60 min at room temperature (RT, 20-25°C).
  3. 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.
  4. Pipette 100 μL of Enzyme Conjugate (HRP-Streptavidin) into each well.
  5. Cover plate with adhesive seal. Shake plate carefully. Incubate 30 min at RT.
  6. 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.
  7. Pipette 100 µL of Ready-to-Use TMB Substrate Solution into each well.
  8. Incubate 10 min at RT. Avoid exposure to direct sunlight.
  9. 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.
  10. 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

Certolizumab Pegol mAb-based ELISA


Product Manual


Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.



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