Vedolizumab mAb-based ELISA Assay

$1,260.00

The Vedolizumab ELISA Assay Kit is an enzyme immunoassay for the specific determination of free Vedolizumab (Entyvio®) in serum and plasma. The Eagle Biosciences Vedolizumab (Entyvio®) ELISA Assay Kit is for research use only and is not intended for diagnostic or therapeutic procedures.

Vedolizumab mAb-based ELISA Assay

Vedolizumab mAb-based ELISA Assay is for Research Use Only

Size: 1 x 96 wells
Sensitivity: 5 ng/mL
Incubation Time: < 2 hours
Sample Type:  Serum, Plasma
Sample Size: 10 µL
Alternative Names: Entyvio Assay


Assay Background

The drug Vedolizumab (trade name Entyvio®) is a humanized immunoglobulin G1 monoclonal antibody that binds exclusively to the lymphocyte integrin α4β7.  Identification of biomarkers might be beneficial for (non-) response and risk factors for adverse drug reactions that might be related to serum drug levels and maintaining the effective concentration in order to potentially avoid some side effects with a reliable method. The specificity of this test system is achieved by using a monoclonal antibody named “IG-19F3” for the coating of the microtiter plate. This antibody is specific for Vedolizumab only and does not cross react with other lymphocyte integrin α4β7 catchers.


STORAGE AND STABILITY OF THE KIT
The kit is shipped at ambient temperature and should be stored at 2-8°C. Keep away from heat or direct sun light. The storage and stability of specimen and prepared reagents is stated in the corresponding chapters. The microtiter strips are stable up to the expiry date of the kit in the broken, but tightly closed bag when stored at 2–8°C.
The usual precautions for venipuncture should be observed. It is important to preserve the chemical integrity of a blood specimen from the moment it is collected until it is assayed. Do not use grossly hemolytic, icteric or grossly lipemic specimens. Samples appearing turbid should be centrifuged before testing to remove any particulate material.


Related Products

Vedolizumab (Entyvio®) Antibodies ELISA Kit
Infliximab (Remicade®) ELISA (mAb-based) Assay Kit
Total Antibodies to Infliximab (Remicade®) ELISA Assay

Additional Information

Assay Principle


This Eagle Biosciences ELISA is based on a Vedolizumab-specific monoclonal antibody (catcher Ab, ImmunoGuide clone19F3). Diluted standards and samples are incubated in the microtiter plate coated with IG-19F3 mAb. After incubation, the wells are washed. A horseradish peroxidase (HRP)-conjugated anti-human IgG antibody is added and binds to the Fc part of Vedolizumab. Following incubation, wells are washed and the bound enzymatic activity is detected by addition of chromogen-substrate. The color developed is proportional to the amount of Vedolizumab 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 100 µL of each 1:10 Diluted Standard, and 1:200 Diluted Samples into the respective wells of the microtiter plate. Bubble formation during the pipetting of standards and samples must be avoided.
  3. Cover the plate with adhesive seal. Shake plate carefully by tapping several times. Incubate the plate on bench top for 60 min at room temperature (RT, 20-25°C) without shaking.
  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) into each well.
  6. Cover plate with adhesive seal. Shake plate carefully by tapping several times.  Incubate the plate on a bench top for 30 min at RT without shaking.
  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 20 min at RT. Avoid exposure to direct sunlight.
  10. Stop the substrate reaction by adding 100 µL of Stop Solution into each well. Color changes from blue to yellow. Briefly mix contents by gently shaking the plate.
  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


Vedolizumab mAb-based ELISA Assay

Manual

Product Manual


Publications

Citations


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  • Bruce E. Sands, Brian G. Feagan, Paul Rutgeerts, Jean-Frédéric Colombel, William J. Sandborn, Richmond Sy, Geert D’Haens et al., Effects of Vedolizumab Induction Therapy for Patients With Crohn’s Disease in Whom Tumor Necrosis Factor Antagonist Treatment Failed, Gastroenterology 2014;147:618–627.
  • Soler D, Chapman T, Yang LL, Wyant T, Egan R, Fedyk ER., The binding specificity and selective antagonism of vedolizumab, an anti-alpha4beta7 integrin therapeutic antibody in development for inflammatory bowel diseases., J Pharmacol Exp Ther. 2009;330(3):864-75.
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    Cohen LB, Nanau RM, Delzor F, Neuman MG. Biologic therapies in inflammatory bowel disease., Transl Res. 2014;163(6):533-56.
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  • Amiot A, Grimaud JC, Peyrin-Biroulet L, Filippi J, Pariente B, et al. Observatory on Efficacy and of Vedolizumab in Patients With Inflammatory Bowel Disease Study Group.; Groupe d’Etude Therapeutique des Affections Inflammatoires du tube Digestif.. Effectiveness and Safety of Vedolizumab Induction Therapy for Patients With Inflammatory Bowel Disease. Clin Gastroenterol Hepatol. 2016;14(11):1593-1601.
  • Loftus EV Jr, Colombel JF, Feagan BG, Vermeire S, Sandborn WJ, Sands BE, Danese S, D’Haens GR, Kaser A, Panaccione R, Rubin DT, Shafran I, McAuliffe M, Kaviya A, Sankoh S, Mody R, Abhyankar B, Smyth M. Long-term Efficacy of Vedolizumab for Ulcerative Colitis. J Crohns Colitis. 2016 Sep 28. pii: jjw177. [Epub ahead of print].
  • Dulai PS, Singh S, Jiang X, Peerani F, Narula N, Chaudrey K, Whitehead D, Hudesman D, Lukin D, Swaminath A, Shmidt E, Wang S, Boland BS, Chang JT, Kane S, Siegel CA, Loftus EV, Sandborn WJ, Sands BE, Colombel JF. The Real-World Effectiveness and Safety of Vedolizumab for Moderate-Severe Crohn’s Disease: Results From the US VICTORY Consortium. Am J Gastroenterol. 2016;111(8):1147-55.
  • Rosario M, Dirks NL, Gastonguay MR, Fasanmade AA, Wyant T, Parikh A, Sandborn WJ, Feagan BG, Reinisch W, Fox I. Population pharmacokinetics-pharmacodynamics of vedolizumab in patients with ulcerative colitis and Crohn’s disease. Aliment Pharmacol Ther. 2015;42(2):188-202.
  • Colombel JF, Sands BE, Rutgeerts P,Sandborn W, Danese S, D’Haens G, Panaccione R, Loftus EV Jr, Sankoh S, Fox I, Parikh A, Milch C, Abhyankar B, Feagan BG. The safety of vedolizumab for ulcerative colitis and Crohn’s disease. Gut. 2016 Feb 18. pii: gutjnl-2015-311079. doi: 10.1136/gutjnl-2015-311079. (Epub ahead of print)
  • Sandborn WJ, Feagan BG, Rutgeerts P, Hanauer S, Colombel JF, Sands BE, Lukas M, Fedorak RN, Lee S, Bressler B, Fox I, Rosario M, Sankoh S, et al.; GEMINI 2 Study Group. Vedolizumab as induction and maintenance therapy for Crohn’s disease. N Engl J Med. 2013;369(8):711-21.
  • Fedyk ER, Wyant T, Yang LL, Csizmadia V, Burke K, Yang H, Kadambi VJ. Exclusive antagonism of the α4 β7 integrin by vedolizumab confirms the gut-selectivity of this pathway in primates. Inflamm Bowel Dis. 2012;18(11):2107-19.
  • Feagan BG, Greenberg GR, Wild G, Fedorak RN, Paré P, McDonald JW, Cohen A, Bitton A, Baker J, Dubé R, Landau SB, Vandervoort MK, Parikh A. Treatment of active Crohn’s disease with MLN0002, a humanized antibody to the alpha4beta7 integrin. Clin Gastroenterol Hepatol. 2008;6(12):1370-7.
  • Yajnik V, Khan N, Dubinsky M, Axler J, James A, Abhyankar B, Lasch K. Efficacy and Safety of Vedolizumab in Ulcerative Colitis and Crohn’s Disease Patients Stratified by Age. Adv Ther. 2017;34(2):542-559.