Adalimumab (Humira®) ELISA Assay Kit

$895.00

The Eagle Biosciences Adalimumab ELISA Assay Kit is an enzyme immunoassay for the quantitative determination of free Adalimumab 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 Entanercept, Infliximab, and Golimumab.  The Adalimumab ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Adalimumab (Humira®) ELISA Assay Kit

For Research Use Only

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

Controls Included


Additional Information

Assay Background

The drug Adalimumab (trade name Humira®) is a recombinant human IgG1 monoclonal antibody specific for human tumor necrosis factor alpha (TNF-α). Adalimumab was created using phage display technology resulting in an antibody with human derived heavy and light chain variable regions and human IgG1-Κ constant regions. Adalimumab is produced by recombinant DNA technology in a mammalian cell expression system and it consists of 1330 amino acids and has a molecular weight of approximately 148 kilodaltons. Adalimumab binds specifically to (TNF-α)) and blocks its interaction with the p55 and p75 cell surface TNF receptors. Adalimumab also lyses surface TNF expressing cells in vitro in the presence of complement.

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 Adalimumab in order to potentially avoid some side effects with a reliable method might be beneficial.

Assay Principle

This Eagle Biosciences Adalimumab 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 Adalimumab 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 Adalimumab 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

Manual

Product Manual


Publications

Citations

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Takahashi H, Tsuji H, Ishida-Yamamoto A, Iizuka H. Plasma trough levels of adalimumab and infliximab in terms of clinical efficacy during the treatment of psoriasis. J Dermatol. 2013;40(1):39-42.

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Oh K, Ito S, Unno M, Kobayashi D, Azuma C, Abe A, Otani H, Ishikawa H, Nakazono K, Narita I, Murasawa A. The rate of decrease in the disease activity of rheumatoid arthritis during treatment with adalimumab depends on the dose of methotrexate. Intern Med. 2015;54(9):1035-41.

Chen DY, Chen YM, Hsieh TY, Hung WT, Hsieh CW, Chen HH, Tang KT, Lan JL. Drug trough levels predict therapeutic responses to dose reduction of adalimumab for rheumatoid arthritis patients during 24 weeks of follow-up. Rheumatology (Oxford). 2015 Aug 31. pii: kev298.

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Eng GP, Bendtzen K, Bliddal H, Stoltenberg M, Szkudlarek M, Fana V, Lindegaard HM, Omerovic E, Højgaard P, Jensen EK, Bouchelouche PN. Antibodies to infliximab and adalimumab in patients with rheumatoid arthritis in clinical remission: a cross-sectional study. Arthritis. 2015;2015:784825. doi: 10.1155/2015/784825.

Bodini G, Savarino V, Peyrin-Biroulet L, de Cassan C, Dulbecco P, Baldissarro I, Fazio V, Giambruno E, Savarino E. Low serum trough levels are associated with post-surgical recurrence in Crohn’s disease patients undergoing prophylaxis with adalimumab. Dig Liver Dis. 2014;46(11):1043-

Tanida S, Mizoshita T, Nishie H, Ozeki K, Katano T, Kubota E, Kataoka H, Kamiya T, Joh T. Combination Therapy With Adalimumab Plus Intensive Granulocyte and Monocyte Adsorptive Apheresis in Patients With Refractory Ulcerative Colitis. J Clin Med Res. 2015;7(11):884-9.

Sohn IW, Kim ST, Kim B, Lee HJ, Park SJ, Hong SP, Kim TI, Kim WH, Cheon JH. Efficacy of Adalimumab in Korean Patients with Crohn’s Disease. Gut Liver. 2015 Oct 19. doi: 10.5009/gnl15165.

Fox JD, Baquerizo-Nole KL, Keegan BR, Macquhae F, Escandon J, Espinosa A, Perez C, Romanelli P, Kirsner RS. Adalimumab treatment leads to reduction of tissue tumor necrosis factor-alpha correlated with venous leg ulcer improvement: a pilot study. Int Wound J. 2015 Sep 24. doi: 10.1111/iwj.12497.

Bálint A, Farkas K, Palatka K, Lakner L, Miheller P, Rácz I, Hegede G, Vincze Á, Horváth G, Szabó A, Nagy F, Szepes Z, Gábor Z, Zsigmond F, Zsóri Á, Juhász M, Csontos Á, Szűcs M, Bor R, Milassin Á, Rutka M, Molnár T. Efficacy and Safety of Adalimumab in Ulcerative Colitis Refractory to Conventional Therapy in Routine Clinical Practice. J Crohns Colitis. 2015 Sep 20. pii: jjv169.

Migliore A, Bizzi E, Egan CG, Bernardi M, Petrella L. Efficacy of biological agents administered as monotherapy in rheumatoid arthritis: a Bayesian mixed-treatment comparison analysis. Ther Clin Risk Manag. 2015;11:1325-35.

Sagami S, Ueno Y, Tanaka S, Nagai K, Hayashi R, Chayama K. Successful Use of Adalimumab for Treating Pyoderma Gangrenosum with Ulcerative Colitis under Corticosteroid-tapering Conditions. Intern Med. 2015;54(17):2167-72.

Thomas SS, Borazan N, Barroso N, Duan L, Taroumian S, Kretzmann B, Bardales R, Elashoff D, Vangala S, Furst DE. Comparative Immunogenicity of TNF Inhibitors: Impact on Clinical Efficacy and Tolerability in the Management of Autoimmune Diseases. A Systematic Review and Meta-Analysis. BioDrugs. 2015 Aug;29(4):241-58.

Otsuka A, Morita M, Yamada H. Clinical characteristics of Japanese patients with axial spondyloarthritis, and short-term efficacy of adalimumab. J Orthop Sci. 2015 Aug 6.

Real-Fernández F, Cimaz R, Rossi G, Simonini G, Giani T, Pagnini I, Papini AM, Rovero P. Surface plasmon resonance-based methodology for anti-adalimumab antibody identification and kinetic characterization. Anal Bioanal Chem. 2015;407(24):7477-85.

Arstikyte I, Kapleryte G, Butrimiene I, Venalis A. Influence of Immunogenicity on the Efficacy of Long-Term Treatment with TNF α Blockers in Rheumatoid Arthritis and Spondyloarthritis Patients. Biomed Res Int. 2015;2015:604872. doi: 10.1155/2015/604872. Epub 2015 Apr 27.

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