Etanercept (Enbrel®) Antibodies ELISA Assay Kit

$895.00

The Eagle Biosciences Antibody to Etanercept is an enzyme immunoassay for the semi-quantitative determination of free antibodies to Etanercept in serum and plasma samples. The Antibody to Etanercept ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

Etanercept (Enbrel®) Antibodies ELISA Assay Kit

For Research Use Only

Size: 1 x 96 wells
Sensitivity: Cut-off
Incubation Time: 2.5 hours
Sample Type:  Serum, Plasma
Sample Size: 10 µL

Additional Information

Assay Background

The drug Etanercept (trade name Enbrel®) is a dimeric fusion protein consisting of the extracellular ligand-binding portion of the human 75 kilodalton (p75) tumor necrosis factor receptor (TNFR) linked to the Fc portion of human IgG1. One of the major concerns, despite of its wide usage, is the potential development of anti-drug antibodies (ADA) which in turn may interfere with the drug efficacy as mainly judged by observing the relapse of signs and symptoms of disease and necessitate dose-escalation or potentially ending up the treatment.

The ImmunoGuide Antibody to Etanercept ELISA Kit can be used for the measurement of free antibodies against this drug. It does not detect such antibodies which already are bound to the drug.

Assay Principle

This Eagle Biosciences ImmunoGuide anti-drug antibody(ies) (ADA) kit is a bridging type ELISA for the determination of free antibodies against the drug Etanercept in serum and plasma samples. During the first incubation period, ADA in serum or plasma samples are captured by the drug coated on the microtiter wells. After washing away the unbound components from samples, a peroxidase-labelled drug conjugate is added and then incubated. ADA, if present in sample, will make a bridge, with its identical Fab arms, between the drug coated on the well and the other drug molecule labelled with peroxidase. After a second washing step, the bound enzymatic activity is detected by addition of tetramethylbenzidine (TMB) chromogen-substrate. Finally, the reaction is terminated with stop solution. The positive reaction is expected to be related to the presence of ADA in the sample.

Assay Procedure

  1. Pipette 100 µL of Assay Buffer into each of the wells to be used.
  2. Pipette 50 µL of each Ready-to-Use Negative Control, Ready-to-Use Positive Control, and 1:10 Diluted Samples (as described in section 10.2) 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-drug) into each well.
  6. Cover plate with adhesive seal. Shake plate carefully. Incubate 60 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.

Manual

Product Manual


Publications

Citations

  • Cheong CU, Chang CP, Chao CM, Cheng BC, Yang CZ, Chio CC. Etanercept attenuates traumatic brain injury in rats by reducing brain TNF- α contents and by stimulating newly formed neurogenesis. Mediators Inflamm. 2013;2013:620837. doi: 10.1155/2013/620837.
  • Jung YS, Park W, Na K. Temperature-modulated noncovalent interaction controllable complex for the long-term delivery of etanercept to treat rheumatoid arthritis. J Control Release. 2013 Oct 28;171(2):143-51.
  • Erdemli Ö, Özen S, Keskin D, Usanmaz A, Batu ED, Atilla B, Tezcaner A. In vitro evaluation of effects of sustained anti-TNF release from MPEG-PCL-MPEG and PCL microspheres on human rheumatoid arthritis synoviocytes. J Biomater Appl. 2014 Oct;29(4):524-42.
  • Chen DY, Chen YM. Response to: ‘Towards optimal cut-off trough levels of adalimumab and etanercept for a good therapeutic response in rheumatoid arthritis. Results of the INMUNOREMAR study’ by Sanmarti et al. Ann Rheum Dis. 2015;74(8):e43. doi: 10.1136/annrheumdis-2015-207543.
  • Jani M, Chinoy H, Warren RB, Griffiths CE, Plant D, Fu B, Morgan AW, Wilson AG, Isaacs JD, Hyrich K, Barton A; Biologics in Rheumatoid Arthritis Genetics and Genomics Study Syndicate Collaborators. Clinical utility of random anti-tumor necrosis factor drug-level testing and measurement of antidrug antibodies on the long-term treatment response in rheumatoid arthritis. Arthritis Rheumatol. 2015 May;67(8):2011-9.
  • Kneepkens EL, Krieckaert CL, van der Kleij D, Nurmohamed MT, van der Horst-Bruinsma IE, Rispens T, Wolbink GJ. Lower etanercept levels are associated with high disease activity in ankylosing spondylitis patients at 24 weeks of follow-up. Ann Rheum Dis. 2015 Oct;74(10):1825-9.
  • Chen DY, Chen YM, Tsai WC, Tseng JC, Chen YH, Hsieh CW, Hung WT, Lan JL. Significant associations of antidrug antibody levels with serum drug trough levels and therapeutic response of adalimumab and etanercept treatment in rheumatoid arthritis. Ann Rheum Dis. 2015 Mar;74(3):e16. doi: 10.1136/annrheumdis-2013-203893.
  • Garcês S, Antunes M, Benito-Garcia E, da Silva JC, Aarden L, Demengeot J. A preliminary algorithm introducing immunogenicity assessment in the management of patients with RA receiving tumour necrosis factor inhibitor therapies. Ann Rheum Dis. 2014 Jun;73(6):1138-43.
  • Mahil SK, Arkir Z, Richards G, Lewis CM, Barker JN, Smith CH. Predicting treatment response in psoriasis using serum levels of adalimumab and etanercept: a single-centre, cohort study. Br J Dermatol. 2013 Aug;169(2):306-13.
  • Sivamani RK, Goodarzi H, Garcia MS, Raychaudhuri SP, Wehrli LN, Ono Y, Maverakis E. Biologic therapies in the treatment of psoriasis: a comprehensive evidence-based basic science and clinical review and a practical guide to tuberculosis monitoring. Clin Rev Allergy Immunol. 2013;44(2):121-40.
  • Hutmacher MM, Nestorov I, Ludden T, Zitnik R, Banfield C. Modeling the exposure-response relationship of etanercept in the treatment of patients with chronic moderate to severe plaque psoriasis. J Clin Pharmacol. 2007;47(2):238-48.
  • Yim DS, Zhou H, Buckwalter M, Nestorov I, Peck CC, Lee H. Population pharmacokinetic analysis and simulation of the time-concentration profile of etanercept in pediatric patients with juvenile rheumatoid arthritis. J Clin Pharmacol. 2005 Mar;45(3):246-56.
  • Andrick BJ, Schwab AI, Cauley B, O’Donnell LA, Meng WS. Predicting Hemagglutinin MHC-II Ligand Analogues in Anti-TNFα Biologics: Implications for Immunogenicity of Pharmaceutical Proteins. PLoS One. 2015 Aug 13;10(8):e0135451. doi: 10.1371/journal.pone.0135451
  • Emery P, Vencovský J, Sylwestrzak A, Leszczyński P, Porawska W, Baranauskaite A, Tseluyko V, Zhdan VM, Stasiuk B, Milasiene R, Barrera Rodriguez AA, Cheong SY, Ghil J. A phase III randomised, double-blind, parallel-group study comparing SB4 with etanercept reference product in patients with active rheumatoid arthritis despite methotrexate therapy. Ann Rheum Dis. 2015 Jul 6. pii: annrheumdis-2015-207588. doi: 10.1136/annrheumdis-2015-207588.
  • 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.
  • Baldo BA. Chimeric fusion proteins used for therapy: indications, mechanisms, and safety. Drug Saf. 2015;38(5):455-79.
  • Deehan M, Garcês S, Kramer D, Baker MP, Rat D, Roettger Y, Kromminga A Managing unwanted immunogenicity of biologicals. Autoimmun Rev. 2015;14(7):569-74.
  • Senabre-Gallego JM, Santos-Ramírez C, Santos-Soler G, Salas-Heredia E, Sánchez-Barrioluengo M, Barber X, Rosas J; AIRE-MB group. Long-term safety and efficacy of etanercept in the treatment of ankylosing spondylitis. Patient Prefer Adherence. 2013;7:961-72.
  • Hunt L, Emery P. Etanercept in the treatment of rheumatoid arthritis. Expert Opin Biol Ther. 2013;13(10):1441-50.
  • Jamnitski A, Bartelds GM, Nurmohamed MT, van Schouwenburg PA, van Schaardenburg D, Stapel SO, Dijkmans BA, Aarden L, Wolbink GJ. The presence or absence of antibodies to infliximab or adalimumab determines the outcome of switching to etanercept. Ann Rheum Dis. 2011;70(2):284-8