Certolizumab Pegol mAb-based ELISA

$1,415.00

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

Documents

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.

Publications

Citations


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  • Loftus EV Jr, Colombel JF, Schreiber S, Randall CW, Regueiro M, Ali T, Arendt C, Coarse J, Spearman M, Kosutic G. Safety of Long-Term Treatment With Certolizumab Pegol in Patients with Crohn’s Disease, Based on a Pooled Analysis of Data From Clinical Trials. Clin Gastroenterol Hepatol. 2016 Jul 24. pii: S1542-3565(16)30440-2. doi: 10.1016/j.cgh.2016.07.019.
  • Lee SD, Rubin DT, Sandborn WJ, Randall C, Younes Z, Schreiber S, Schwartz DA, Burakoff R, Binion D, Dassopoulos T, Arsenescu R, Gutierrez A, Scherl E, Kayhan C, Hasan I, Kosutic G, Spearman M, Sen D, Coarse J, Hanauer S. Reinduction with Certolizumab Pegol in Patients with Crohn’s Disease Experiencing Disease Exacerbation: 7-Year Data from the PRECiSE 4 Study. Inflamm Bowel Dis. 2016; 22(8): 1870-80.
  • Cassinotti A, Ardizzone S, Porro GB. Certolizumab pegol: an evidence-based review of its place in the treatment of Crohn’s disease. Core Evid. 2008; 2(3): 209-29.
  • Porter C, Armstrong-Fisher S, Kopotsha T, Smith B, Baker T, Kevorkian L, Nesbitt A. Certolizumab pegol does not bind the neonatal Fc receptor (FcRn): Consequences for FcRn-mediated in vitro transcytosis and ex vivo human placental transfer. J Reprod Immunol. 2016; 116: 7-12.
  • Lee YH, Bae SC. Efficacy and safety of methotrexate plus certolizumab pegol or placebo in active rheumatoid arthritis : Meta-analysis of randomized controlled trials. Z Rheumatol 2016 Jun 16. DOI10.1007/s00393-016-0133-z
  • Rudwaleit M, Rosenbaum JT, Landewé R, Marzo-Ortega H, Sieper J, van der Heijde D, Davies O, Bartz H, Hoepken B, Nurminen T, Deodhar A. Observed Incidence of Uveitis Following Certolizumab Pegol Treatment in Patients With Axial Spondyloarthritis. Arthritis Care Res (Hoboken). 2016; 68(6): 838-44.
  • Pope J, Bingham CO 3rd, Fleischmann RM, Dougados M, Massarotti EM, Wollenhaupt J, Duncan B, Coteur G, Weinblatt ME. Impact of certolizumab pegol on patient-reported outcomes in rheumatoid arthritis and correlation with clinical measures of disease activity. Arthritis Res Ther. 2015; 17: 343.
  • Weinblatt ME, Fleischmann R, van Vollenhoven RF, Emery P, Huizinga TW, Cutolo M, van der Heijde D, Duncan B, Davies O, Luijtens K, Dougados M. Twenty-eight-week results from the REALISTIC phase IIIb randomized trial: efficacy, safety and predictability of response to certolizumab pegol in a diverse rheumatoid arthritis population. Arthritis Res Ther. 2015; 17(1): 325.
  • Mease P, Deodhar A, Fleischmann R, Wollenhaupt J, Gladman D, Leszczyński P, Vitek P, Turkiewicz A, Khraishi M, FitzGerald O, Landewé R, de Longueville M, Hoepken B, Peterson L, van der Heijde D. Effect of certolizumab pegol over 96 weeks in patients with psoriatic arthritis with and without prior antitumour necrosis factor exposure. RMD Open. 2015 Jun 25;1(1):e000119. doi: 10.1136/rmdopen-2015-000119. eCollection 2015
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